Method for production of CTLA-4 binding antibody, antibody obtained by the use of this method, pharmaceutical composition, cell line, isolated nucleic acid, host cell, transgenic mammal or plant and the use of human monoclonal CTLA-4 binding antibody, fragment thereof or pharmaceutical composition and their use in combination with GM-CSF

The present invention relates to a monoclonal antibody that binds to a cytotoxic T cell antigen (CTLA-4) or an antigen-binding fragment thereof, and a method for producing the antibody. These antibodies may find application in particular in chronic diseases where repeated administration of antibodies is required.

This application claims priority to Provisional Patent Application Serial No. US 60 / 113,647 filed December 23, 1998, the contents of which are hereby incorporated by reference.

Regulation of the immune response in patients would enable the treatment of many human diseases which would have specific effects so rarely seen with conventional drugs. It would be possible to both raise and lower the immune system response. The role of T cells and B cells in regulating the immune response has been extensively studied and described. These studies show that in many cases the role of T cells seems to be particularly important in the prevention and treatment of diseases.

T cells have very complex systems for controlling their own interactions. Numerous receptors and soluble factors are used in the processes of interaction between T cells. Thus, what affects one particular signal can alter the overall immune response and depend on many specific factors, receptors and counter receptors that are involved in a given pathway. Response lowering pathways are as important as those required for activation. One mechanism for preventing an immune response to a particular antigen is through the process of "education" in the thymus leading to T-cell tolerance. Other mechanisms such as the secretion of suppressive cytokines are also known.

Activation of T cells requires not only stimulation through the antigen receptor (T cell receptor (TOR)), but also additional signals through costimulatory surface molecules such as CD28. The ligands for CD28 are white B7-1 (CD80) and B7-2 (CD86), which are expressed on antigen presenting cells such as dendritic cells, activated B cells, or monocytes that interact with CD28 or CTLA-4 T cells to providing a costimulatory signal. The role of costimulatory signaling has been investigated in experimental allergic encephaloyelitis (EAE) by Perrin et al., Immunol Res 14: 189-99 (1995). EAE is an autoimmune disease induced by Th1 cells directed against myelin antigens that provides an in vivo model to study the role of B7 co-stimulation in inducing pathological immune responses. Using a soluble fusion protein as a ligand for the B7 receptor as well as monoclonal antibodies specific for CD80 or CD86, Perrin et al. Demonstrated that B7 costimulation plays an important role in EAE in determining the clinical effect of the disease.

The interaction between B7 and CD28 is one of several costimulatory signal transduction pathways that appear sufficient to engage the maturation and proliferation of antigen-specific T cells. The lack of costimulation, and the associated insufficient production of IL-2, prevents the subsequent proliferation of T cells and induces a state of non-reactivity called "anergy. Many viruses and tumors can block the activation and proliferation of T cells, leading to insufficient activity or inactivity of the host's immune system against infected or transformed cells. Among possible T cell dysfunctions, anergy may be at least in part responsible for the host's inability to eliminate pathogenic or neoplastic cells.

The use of the B7 protein in mediating anti-tumor immunity is described in Chen et al., Cell 71: 1093-1102 (1992) and Townsend and Allison, Science 259: 368 (1993). Schwartz's review publication, Cell 71: 1065 (1992) describes the role of CD28, CTLA-4 and B7 in IL-2 production and immunotherapy. Harding et al., Nature 356: 607-609 (1994) have shown that CD28-mediated signaling stimulates murine T cells and prevents the induction of anergy in T cell clones. See also U.S. Patent Nos. 5,434,131,5,770,197 and 5,773,253 and International Patent Applications Nos. WO 93/00431, WO 95/01994, WO 95/03408, WO 95/24217 and WO 95/33770.

It is clear from the above considerations that T cells require two types of antigen presenting (APC) signals in order to be activated and subsequently differentiated to exhibit effector functions. First, there is an antigen-specific signal produced by the interaction between the TCR on T cells and the junction

PL 214 003 Β1 MHC molecules presenting the peptides on the APC. Second, there is an antigen-independent signal that is mediated by CD28 interaction with members of the B7 family (B7-1 (CD80) or B7-2 (CD86)). The exact positioning of CTLA-4 in the immune response processes was initially unclear. Murine CTLA-4 was first identified and cloned by Brunet et al., Nature 328: 267-270 (1987), in the search for molecules that are preferentially expressed on cytotoxic T cells. Human CTLA-4 was identified and cloned soon later by Dariavach et al. Eur. J. Immunol. 18: 1901-1905 (1988). Mouse and human CTLA-4 molecules share approximately 76% full sequence homology and sequence identity in the cytoplasmic domains (Dariavach et al. Eur. J. Immunol. 18: 1901-1905 (1988)). CTLA-4 is a member of the immunoglobulin (Ig) superfamily of proteins. The Ig superfamily is a group of proteins that share the structural properties of the variable (V) or constant (C) domains of Ig molecules. Members of this Ig superfamily include, but are not limited to, immunoglobulins themselves, major histocompatibility complex (MHC) molecules (i.e., MHC classes I and II), and TCR molecules.

In 1991, Linsley et al., J. Exp. Med. 174: 561-569 (1991), proposed that CTLA-4 is the second receptor for B7. Similarly, Harper et al., J Immunol 147: 1037-44 (1991) have shown that CTLA-4 and CD28 molecules are closely related in mouse and human with respect to sequence, mRNA expression, gene structure, and chromosomal location. See also Balzano et al., Int J Cancer Suppl 7: 28-32 (1992). Further evidence for this role came from functional studies. For example, Lenschow et al., Science 257: 789-79 (1992) showed that CTLA-4-Ig induced long-term survival of pancreatic islet grafts. Freeman et al., Science 262: 907-909 (1993) investigated the role of CTLA-4 in B7 deficient mice. The study of ligands for CTLA-4 is described in Lenschow et al., PNAS 90: 11054-11058 (1993). Linsley et al., Science 257: 792-795 (1992) describe in vivo immunosuppression induced by a soluble form of CTLA-4. Linsley et al., J Exp Med 176: 1595-604 (1992) prepared antibodies that bound CTLA-4 and that did not cross-react with CD28 and concluded that CTLA-4 is co-expressed with CD28 on activated T cells and cooperatively regulates T cell adhesion and activation by B7. Kuchroo et al., Cell 80: 707-18 (1995) showed that costimulatory molecules B7-1 and B7-2 activated Th1 / Th2 development pathways in a differentiated manner. Yiqun et al., Int Immunol 8: 37-44 (1996) demonstrated that there are differential requirements for costimulatory signals from members of the B7 family for resting memory T cells versus recently activated memory T cells for soluble booster antigens. See also de Boer et al. Eur J Immunol 23: 3120-5 (1993).

Several groups have proposed alternative or distinct receptor / ligand interactions for CTLA-4 compared to CD28, and have even proposed a third B-7 complex that is recognized by BB1 antibodies. See, for example, Hathcock et al. Science 262: 905-7 (1993), Freeman et al. Science 262: 907-9 (1993), Freeman et al. J Exp Med 178: 2185-92 (1993), Lenschow et al. Proc Natl Acad Sci USA 90: 11054-8 (1993), Razi-Wolfi et al. Proc Natl Acad Sci USA 90: 11182-6 (1993) and Boussiotis et al. Proc Natl Acad Sci USA 90: 11059-63 (1993). But in Freeman et al. J Immunol 161: 2708-15 (1998) the discovery that antibody BB1 binds to a molecule that is identical to the surface form of CD74, postulating that BB1 mAb binds to a protein other than B7-1. and this epitope is also present on the B7-1 protein. Therefore, it became necessary to reconsider the use of BB1 mAb in the analysis of CD80 expression and function.

In studies from 1993-1995, scientists made further efforts to determine the role that CTLA-4 plays in T-cell stimulation. First, by using monoclonal antibodies against CTLA-4, Walunas et al., Immunity 1: 405-13 (1994) provided evidence that CTLA-4 may act as a negative regulator of T cell activation. Then, Waterhouse et al., Science 270: 985-988 (1995) showed that CTLA-4 deficient mice accumulated in lymph nodes and spleens T precursor cells (blastocytes, T blasts) with elevated levels of activation markers. Precursor cells infiltrated liver, heart, lung and pancreatic tissue, serum immunoglobulin levels were increased, and T cells proliferated spontaneously and strongly upon T cell receptor stimulation, however they were susceptible to cell death induced by Fas receptor cross-linking and by gamma radiation . Waterhouse et al. Concluded that CTLA-4 acts as a negative regulator of T cell activation and is important in the control of lymphocyte homeostasis. As a commentary on this, Allison and Krummel, Science 270: 932-933 (1995) discussed the work of Waterhouse et al. As showing that CTLA-4 either acts to lower T cell response or plays a role in inhibiting signal transduction in T cell activation and development. Tivol et al., Immunity 3: 541-7 (1995) also obtained CTLA-4 deficient mice and showed that such

PL 214 003 Β1 mice rapidly develop disease associated with the proliferation of white blood cells with multi-organ lymphocytic infiltration and tissue destruction with particularly acute myocarditis and pancreatitis. They concluded that CTLA-4 plays a key role in reducing T cell activation and maintaining immune homeostasis. Moreover, Krummel and Allison, J Exp Med 182: 459-65 (1995) further elucidate that CD28 and CTLA-4 exert an opposite effect on the T cell response to stimulation. They produced an anti-CTLA-4 antibody and tested the effect of its binding to CTLA-4 in a system using highly purified T cells. In their report, they showed that the presence of low levels of B7-2 on freshly explanted T cells may partially inhibit T cell proliferation, and that inhibition was mediated by interactions with CTLA-4. Cross-linking of CTLA-4 simultaneously with TCR and CD28 strongly inhibited the proliferation and secretion of IL-2 by T cells. Finally, these results showed that CD28 and CTLA-4 provide opposing signals that appear to be integrated by T cells in determining antigen responses . Thus, they recognized that the result of T cell receptor stimulation is regulated by CD28 costimulatory signals as well as inhibitory signals delivered by CTLA-4. See also Krummel et al. Int Immunol 8: 519-23 (1996) and US Patent No. 5,811,097 and International Patent Application No. WO 97/20574.

Various additional experiments were performed to explain the above function of CTLA-4 in more detail. For example, Walunas et al., J Exp Med 183: 2541-50 (1996), by using an anti-CTLA-4 antibody, suggested that CTLA-4 signaling does not regulate cell survival or IL-2 response, but inhibits CD28 dependent IL-2 production. Moreover, Perrin et al. J Immunol 157: 1333-6 (1996) showed that anti-CTLA-4 antibodies in experimental encephalomyelitis (EAE) aggravated disease and increased mortality. Disease worsening was accompanied by increased production of TNF-alpha, IFN-gamma and IL-2 cytokines causing encephalomyelitis. Thus, they found that CTLA-4 regulates the intensity of the immune response in EAE, suppressing inflammatory cytokine production and the clinical symptoms of the disease. See also Hurwitz et al. J Neuroimmunol 73: 57-62 (1997) and Cepero et al. J Exp Med 188: 199-204 (1998) (a hairpin type anti-CTLA-4 ribozyme that specifically abolishes CTLA-4 expression after transfer to mouse T-cell model).

In addition, Blair et al., J Immunol 160: 12-5 (1998) investigated the functional effects of a panel of monoclonal antibodies (mAbs) against CTLA-4 on resting human CD4 + T cells. Their results showed that some CTLA-4 mAbs could inhibit the proliferative responses of resting CD4 + cells and the cell cycle transition from GO to G1. The inhibitory effects of CTLA-4 were seen within 4 hours, while expression of CTLA-4 on the cell surface remained undetectable. However, the other mAbs against CTLA-4 showed no detectable inhibitory effects, indicating that binding of the mAb to CTLA-4 alone is not sufficient to mediate a reduction in the level of T-cell response. Interestingly, once IL-2 production is stopped, the inhibitory anti-inhibitory -CTLA-4 allowed the induction and expression of the cell survival gene, bcl-X (L). Consistent with this observation, cells remained viable and no apoptosis was observed when combined with CTLA-4.

In view of the anergy, Perez et al., Immunity 6: 411-7 (1997) showed that the induction of T-cell anergy was counteracted by CTLA-4 blocking, and concluded that the outcome of antigen recognition by T cells was conditioned by CD28 or CTLA-4 interaction. on T cells with B7 molecules. Also, Van Parijs et al., J Exp Med 186: 1119-28 (1997) investigated the role of interleukin 12 and costimulators in T cell anergy in vivo and found that by inhibiting the involvement of CTLA-4 in inducing anergy, cell proliferation was blocked. T and there was no complete Th1 differentiation. However, T cells exposed to tolerant antigens in the presence of both IL-12 and anti-CTLA-4 antibodies were not anergic and behaved identically to T cells that were contacted with the immunogenic antigen. These results suggest that two processes are involved in inducing anergy in vivo: participation of CTLA-4, which blocks the ability of T cells to proliferate, and the absence of the prototypical inflammatory cytokine, IL-12, which prevents T cells from differentiating into Th1 effector cells. The combination of IL-12 and anti-CTLA-4 antibodies was sufficient to convert a stimulus that is normally tolerated into an immunogenic stimulus.

In connection with infections, McCoy et al., J Exp Med 186: 183-7 (1997) showed that anti-CTLA-4 antibodies greatly enhanced and accelerated the T cell immune response to Nippostrongylus brasiliensis, resulting in a significant reduction in adult worm numbers. and early termination of the parasite's egg production. See also Murphy et al. J. Immunol. 161: 4153-4160 (1998) (Leishmania donovani).

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In connection with cancer, Kwon et al., PNAS USA 94: 8099-103 (1997) developed a syngeneic murine prostate cancer model and investigated two distinct treatments to elicit an anti-prostate cancer response through enhanced T-cell costimulation: (i) providing direct costimulation by prostate cancer cells transduced to express B7.1 ligand and (ii) in vivo antibody-mediated blockade of CTLA-4 T cells, preventing a reduction in T cell responses. In vivo antibody-mediated blockade of CTLA-4 has been shown to enhance immune responses against prostate cancer. Also, Yang et al. Cancer Res 57: 4036-41 (1997) investigated whether blockade of CTLA-4 function led to enhancement of anti-tumor T cell responses at various stages of tumor growth. Based on the in vitro and in vivo results, they found that blockage of CTLA-4 in people with cancer enhanced their ability to produce anti-tumor T-cell responses, but the occurrence of such an enhanced effect in their model was limited to the early stages of tumor development. Moreover, Hurwitz et al. Proc Natl Acad Sci USA 95: 10067-71 (1998) investigated the generation of T cell mediated anti-tumor responses and found that it depends on T cell receptor involvement by the major histocompatibility complex / antigen as well as on B7 binding of CD28.

Certain tumors, such as SM1 breast cancer, have been refractory to anti-CTLA-4 immunotherapy. Thus, when using the combination of CTLA-4 blockade and SM1 cell vaccine expressing granulocyte-macrophage colony stimulating factor, regressions of parental SM1 tumor cells were observed despite the ineffectiveness of either treatment alone. Combination therapy led to long-lasting immunity to SM1 and was dependent on both CD4 (+) and CD8 (+) T cells. These findings suggested that CTLA-4 blockade acts at the level of host-derived antigen presenting cells.

With regard to diabetes mellitus, Luhder et al., J Exp Med 187: 427-32 (1998) injected anti-CTLA-4 mAb from a TCR transgenic mouse model of diabetes at various stages of the disease. They found that the involvement of CTLA-4 at the time when potentially diabetic T cells are first activated is a crucial event; if their participation is possible, islet invasion ensues, but remains quite harmless for months. If not, islet inflammation with lymphocytic infiltration is more aggressive and diabetes progresses rapidly.

With respect to vaccine immunization, Horspool et al., J Immunol 160: 2706-14 (1998) found that intact anti-CTLA-4 mAb, but not Fab fragments, inhibited the primary humoral response to pCIA / beta gal without affecting responses. resembling, indicating that CTLA-4 activation inhibited Ab production but not T cell induction. Ligand blockade for CD28 and CTLA-4, CD80 (B7-1) and CD86 (B7-2) showed distinct, non-overlapping functions . Blockade of CD80 with the initial immunization completely abolished the primary and secondary antibody immune responses, whereas blockade of CD86 abolished the primary, but not secondary, responses. Co-blockade of CD80 + CD86 was less effective at abolishing Ab responses than either alone. The enhancement of costimulation by co-injection of B7 expressing plasmids increased CTL responses but not Ab responses and did not indicate a Th1 to Th2 transition. These findings indicate a complex and distinct role for CD28, CTLA-4, CD80 and CD86 in the costimulation of T cells after nucleic acid inoculation.

Regarding allograft rejection, Markees et al., J Clin Invest 101: 2446-55 (1998) found in a mouse skin allograft rejection model that graft acceptance is initially dependent on the presence of INF-gamma, CTLA-4 and CD4 T cells ( +). The addition of anti-CTLA-4 or anti-IFN-gamma mAb to the protocol was accompanied by rapid graft rejection, while the anti-IL-4 mAb had no effect.

In view of the role of CTLA-4 with respect to CD28, Fallarino et al., J Exp Med 188: 205-10 (1998) generated TCR transgenic / recombinase activating gene deficient mice / wild type for CD 28 or with CD28 deficiency which has been immunized with an antigen-expressing tumor. Triggered T cells from both types of mice produced cytokines and proliferated in response to stimulating cells deficient in B7 expression. However, while the CD28 + / + T cell response was enhanced by B7-1 costimulation, the CD28 - / - T cell response was strongly inhibited. This inhibition was reversed by monoclonal antibodies against B7-1 or CTLA-4. Thus, CTLA-4 can potently inhibit T cell activation in the absence of CD28, indicating that TCR-mediated signal antagonism is sufficient to explain the inhibitory effect of CTLA-4. Also, Lin et al., J Exp Med 188: 199-204 (1998) studied

PL 214 003 Β1 cardiac allograft rejection in CD28 deficient mice. H-2 (q) hearts were transplanted into either wild-type allogeneic or CD28 deficient mice (H-2 (b)). Graft rejection was delayed in CD28 deficient mice compared to wild-type mice. Treatment of wild type recipients with immunoglobulin-CTLA-4 (CTLA-4-Ig) or anti-B7-1 plus anti-B7-2 mAb significantly prolonged allograft survival. In contrast, treatment of mice deficient in CD28 with CTLA-4-Ig, anti-B7-1 plus anti-B7-2 mAb or anti-CTLA-4 blocking mAb induced an acceleration of graft rejection. In untreated wild-type mice and in CD28 deficient mice treated with CTLA-4-Ig or anti-CTLA-4 mAb, the increased rate of graft rejection was accompanied by severe infiltration of mononuclear cells and increased levels of IFN-gamma and IL-6 transcripts in the transplanted hearts. Thus, the negative regulatory role of CTLA-4 extends beyond its potential ability to prevent CD28 activation through ligand competition. Even in the absence of CD28, CTLA-4 plays an inhibitory role in regulating allograft rejection.

Work was also undertaken on the further description of CTLA-4 expression. For example, Alegre et al., J Immunol 157: 4762-70 (1996) proposed that surface CTLA-4 internalized rapidly, which may explain the low expression levels essentially detected on the cell surface. They concluded that both CD28 and IL-2 play an important role in increasing CTLA-4 expression. Moreover, the accumulation of CTLA-4 on the cell surface appears to be regulated primarily by its rapid endocytosis. Also, Castan et al. Immunology 90: 265-71 (1997), based on an immunohistological analysis of CTLA-4 expression in situ, suggested that T cells in the proliferative medium that were positive for CTLA-4 should be important for immune regulation. .

Accordingly, in view of the wide application and key role that CTLA-4 appears to play in the immune response mechanism, it would be desirable to generate antibodies to CTLA-4 that can be effectively used in immunotherapy. In addition, it would be desirable to obtain antibodies to CTLA-4 that could be used in chronic diseases where repeated administration of the antibodies is required.

Brief description of the figures

Fig. 1 shows the series of nucleotide and amino acid sequences of the kappa heavy and light chain of immunoglobulin molecules: 4.1.1 (Fig. 1A), 4.8.1 (Fig. 1B), 4.14.3 (Fig. 1C), 6.1.1 (Fig. 1A), 1D), 3.1.1 (Fig. 1E), 4.10.2 (Fig. 1F), 2.1.3 (Fig. 1G), 4.13.1 (Fig. 1H), 11.2.1 (Fig. 11), 11.6 .1 (Fig. 1J), 11.7.1 (Fig. 1K), 12.3.1.1 (Fig. 1L), and 12.9.1.1 (Fig. 1M).

Figure 2 shows the alignment of the predicted heavy chain amino acid sequences from clones 4.1.1, 4.8.1, 4.14.3, 6.1.1, 3.1.1, 4.10.2, 4.13.1, 11.2.1, 11.6.1, 11.7. 1, 12.3.1.1 and 12.9.1.1 and the germline amino acid sequence of DP-50 (3-33). Differences between DP-50 germline sequence and clone sequence are shown in bold. The figure also shows the positions of the CDR1, CDR2 and CDR3 sequences as a shade.

Figure 3 shows the alignment of the predicted amino acid sequence of the heavy chain of clone 2.1.3 and the germline amino acid sequence of DP-65 (4-31). Differences between the DP-65 germline sequence and the clone sequence are shown in bold. In the figure the positions of the CDR1, CDR2 and CDR3 sequences of the antibody are underlined.

Figure 4 shows an alignment of the predicted amino acid sequence of the kappa light chain of clones 4.1.1,4.8.1. 4.14.3, 6.1.1,4.10.2 and 4.13.1 and the germline A27 amino acid sequence. Differences between the A27 germline sequence and the clone sequence are shown in bold. In the figure the positions of the CDR1, CDR2 and CDR3 sequences of the antibody are underlined. The deletions observed in the CDR1 of clones 4.8.1,4.14.3 and 6.1.1 are marked as "0".

Figure 5 shows the alignment of the predicted kappa light chain amino acid sequence of clones 3.1.1, 11.2.1, 11.6.1, and 11.7.1 and the germline 012 amino acid sequence. Differences between the germline 012 sequence and the clone sequence are shown in bold. In the figure the positions of the CDR1, CDR2 and CDR3 sequences of the antibody are underlined.

Figure 6 shows the alignment of the predicted amino acid sequence of the kappa light chain of clone 2.1.3 and the germline A10 / A26 amino acid sequence. Differences between the 012 germline sequence and the clone sequence are shown in bold. In the figure the positions of the CDR1, CDR2 and CDR3 sequences of the antibody are underlined.

Figure 7 shows an alignment of the predicted amino acid sequence of the kappa light chain of clone 12.3.1 and the germline A17 amino acid sequence. Differences between the A17 germline sequence and the clone sequence are shown in bold. In the figure the positions of the CDR1, CDR2 and CDR3 sequences of the antibody are underlined.

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Figure 8 shows an alignment of the predicted amino acid sequence of the kappa light chain of clone 12.9.1 and the germline amino acid sequence of A3 / A19. Differences between the A3 / A19 germline sequence and the clone sequence are shown in bold. In the figure the positions of the CDR1, CDR2 and CDR3 sequences of the antibody are underlined.

Figure 9 is a summary of the N-terminal amino acid sequences generated by direct sequencing of the antibody heavy and light chain proteins.

Figure 10 provides some additional information regarding the characteristics of some of the antibodies disclosed herein. In Fig. 10A, the data for clones 3.1.1, 4.1.1, 4.8.1, 4.10.2, 4.14.3 and 6.1.1 are collected. Concentration data, isoelectric focusing (IEF), SDS-PAGE, size exclusion chromatography (SEC), liquid chromatography / mass spectroscopy (LCMS), mass spectroscopy (MALDI) of the N-terminal light chain sequences are shown. Additional details regarding the IEF are provided in Fig. 10B; relating to SDS-PAGE are shown in 10C; and the SEC of the antibody 4.1.1 on 10D.

Figure 11 shows the expression of B7-1 and B7-2 on Raji cells using anti-CD80-PE and anti-CD86-PE mAbs.

Figure 12 shows the concentration-dependent enhancement of IL-2 production in the T / Raji blastocyte assay induced by blocking anti-CTLA-4 antibodies (BNI3, 4.1.1,4.8.1 and 6.1.1).

Figure 13 shows the concentration-dependent enhancement of IFN-γ production by T / Raji blastocytes induced by blocking anti-CTLA-4 antibodies (BNI3, 4.1.1, 4.8.1 and 6.1.1) (same donor T cells).

Figure 14 shows the mean enhancement of IL-2 production in T cells from 6 donors induced by blocking anti-CTLA-4 antibodies in the T / Raji blastocyst assay.

Figure 15 shows the mean enhancement of IFN-γ production in T cells from 6 donors induced by blocking anti-CTLA-4 antibodies in the T / Raji blastocyte assay.

Fig. 16 shows the enhancement of IL-2 production in hPBMC from 5 donors induced by blocking anti-CTLA-4 mAb measured 72 hours after stimulation with SEA.

Fig. 17 shows the enhancement of IL-2 production in whole blood from 3 donors induced by anti-CTLA-4 blocking mAb measured 72 and 96 hours after stimulation with SEA.

Fig. 18 shows the inhibition of tumor growth by an anti-mouse-CTLA-4 antibody in a murine fibrosarcoma model.

Fig. 19 shows the enhancement of IL-2 production induced by anti-CTLA-4 antibodies (4.1.1 and 11.2.1) in 72 hour T / Raji blastocytes and Superantigen assays (whole blood and peripheral blood mononuclear cells from 6 patients).

Fig. 20 shows the dose-dependent enhancement of IL-2 production induced by anti-CTLA-4 antibodies (4.1.1 and 11.2.1) in a 72 hour T / Raji blast cell assay.

Figure 21 shows the dose-dependent enhancement of IL-2 production induced by anti-CTLA-4 antibodies (4.1.1 and 11.2.1) in a 72 hour Superantigen Assay for whole blood stimulated with 100 ng / ml superantigen.

Fig. 22 depicts a series of additional nucleotide and amino acid sequences of the following anti-CTLA-4 antibody chains: full-length heavy chain 4.1.1 (cDNA sequence 22 (a), genomic sequence 22 (b) and amino acid sequence 22 (c)), full-length non-glycosylated heavy chain 4.1.1 (cDNA sequence 22 (d) and amino acid sequence 22 ( e)), light chain 4.1.1 (cDNA sequence 22 (f) and amino acid sequence 22 (g)), full-length heavy chain 4.8.1 (cDNA sequence 22 (h) and amino acid sequence 22 (i)), light chain 4.8.1 (cDNA 22 (j) and amino acid sequence 22 (k)), full-length heavy chain 6.1.1 (cDNA sequence 22 (1) and amino acid sequence 22 (m)), light chain 6.1.1 (cDNA sequence 22 (n) and amino acid sequence 22 (o)), full-length heavy chain 11.2. 1 (cDNA sequence 22 (p) and amino acid sequence 22 (q)) and light chain 11.2.1 (cDNA sequence 22 (r) and amino acid sequence 22 (s)).

The invention relates to a monoclonal antibody that binds to CTLA-4, or an antigen-binding fragment thereof, comprising a heavy chain variable region at least 85% identical to the heavy chain variable region of SEQ. ID NO: 63, and the light chain variable region is at least 90% identical to the light chain variable region of SEQ. Preferably, the heavy chain variable region is at least 90% identical to the heavy chain variable region of SEQ. ID NO .: 63.

Preferably, the antibody or fragment of the invention binds to CTLA-4 with K_D amounting to 10 '⁹ M or more affinity.

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More preferably, the antibody or fragment of the invention inhibits the binding between CTLA-4 and B7-2 to IC₅₀ of 100 nM or less.

Most preferably, the antibody or fragment of the invention binds to CTLA-4 of K_D amounting to 10 '⁹ M or higher affinity and inhibits the binding between CTLA-4 and B7-2 to IC₅₀ of 100 nM or less.

In another aspect, the invention provides a method of producing an antibody of the invention comprising the steps of:

a) immunizing a non-human mammal with an immunogen comprising CTLA-4, wherein the mammal is capable of expressing human antibodies in its B cells;

b) isolating B cells from a mammal;

c) screening B cells, or cell lines derived therefrom, for antibodies that bind CTLA-4;

d) culturing a cell line expressing antibodies that bind to CTLA-4; and

e) isolating antibodies that bind to CTLA-4.

Detailed Description of the Invention

In accordance with the present invention, complete human monoclonal antibodies to human CTLA-4 are provided. Also provided herein are nucleotide sequences encoding amino acid sequences including the light and heavy chain of immunoglobulin molecules, and in particular the sequences corresponding to the heavy and light chain contiguous sequences from FR1 and CDR1 to CDR3 and FR4. In addition, disclosed herein are antibodies with similar binding properties and antibodies (or other antagonist agents) with similar functional properties to the antibodies described herein. Further disclosed herein are hybridomas expressing such immunoglobulin molecules and monoclonal antibodies.

Definitions

Unless otherwise defined, scientific and technical terms used in connection with the present invention have the meanings commonly understood by one skilled in the art. Further, unless required by context, singular terms shall include pluralities and plural terms shall include the singular. In general, the nomenclature and techniques used in this application for cell and tissue culture, molecular biology, protein chemistry, oligo- and polynucleotides, and hybridization are well known and used in the art. Standard techniques are used to generate recombinant DNA molecules, synthesize oligonucleotides, culture, and transform (e.g., electroporation, lipofection) of tissues. Enzymatic reactions and purification processes were performed according to manufacturers' recommendations, methods commonly known in the art, or as described in this application. The techniques and procedures described below are generally performed according to conventional methods well known in the art as described in many general and more specific publications cited and discussed herein. See, e.g., Sambrook et al. Molecular Cloning: A Laboratory Manual (5th ed. 2, Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY (1989)). The nomenclature, techniques, and laboratory procedures used herein to refer to analytical chemistry, organic synthesis, medicinal chemistry, and pharmacy are well known in the art and commonly used in the art. Standard techniques have been used for chemical syntheses, chemical analyzes, pharmaceutical preparation and delivery, and treatment of patients.

Unless otherwise indicated, the following terms used in this application should be understood to have the following meanings:

The term "isolated polynucleotide" as used in this application means a polynucleotide of genome, cDNA, or synthetic origin, or a combination of the above, which due to its origin (1) is not linked to all or part of the polynucleotide in which the "isolated polynucleotide" resides. in nature, (2) is operably linked to a polynucleotide to which it is not naturally linked, or (3) does not naturally occur as part of a larger sequence.

The term "isolated protein" as used in this application refers to a protein based on cDNA, recombinant RNA, or a protein of synthetic origin or a combination thereof, which due to its origin or source from which it was obtained (1) is not associated with proteins naturally occurring, (2) is free of other proteins from the same source, e.g., free of murine proteins, (3) is expressed by cells of a different species, or (4) is not naturally occurring.

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The term "polypeptide" as used herein means a general term relating to native protein, fragments thereof, or analogs of a polypeptide sequence. Thus, a native protein, portions, and analogs of a polypeptide sequence are species of a generally defined group of polypeptides (genes). Preferably, the peptides may include human immunoglobulin heavy chain molecules and human immunoglobulin light chain kappa molecules as shown in Figure 1. as well as antibody molecules formed by linking immunoglobulin heavy chains with immunoglobulin light chains, such as immunoglobulin kappa light chain molecules, and vice versa, as well as fragments and analogs thereof.

As used herein in relation to an object, the term "naturally occurring" or "occurring in nature" refers to an object that can be found in nature. For example, a naturally occurring polypeptide or polynucleotide sequence that is present in an organism (including viruses) may be extracted from a source in nature, and has not been intentionally altered by man in the laboratory or otherwise.

The term "operably linked" as used in this application refers to the combination of the components of the term such that they can perform as intended. A control sequence "operably linked to a coding sequence is ligated thereto in such a way that expression of the coding sequence is achieved under conditions appropriate to the control sequence.

The term "control sequence" as used in this application refers to polynucleotide sequences necessary to effect the expression and processing of coding sequences to which they are linked. Such control sequences vary with the host organism; in prokaryotes, control sequences typically include a promoter, ribosome binding site, and a transcription termination sequence; in eukaryotes, typically, such control sequences include promoters and transcription termination sequences. The term "control sequences" is intended to include at least all components whose presence is necessary for expression and processing, and may also include additional components whose presence is advantageous, such as, for example, leader sequences and fusion partner sequences.

The term "polynucleotide" as used in this application refers to a polymeric form of nucleotides of at least 10 bases in length, both ribonucleotides and deoxyribonucleotides or modified forms of either type of nucleotide. The term includes single and double stranded forms of DNA.

The term "oligonucleotide" as used in this application refers to naturally occurring and modified nucleotides joined by naturally occurring and non-naturally occurring oligonucleotide linkages. Oligonucleotides are a subset of polynucleotides that are typically 200 bases or less. Preferably the oligonucleotides are 10 to 60 bases in length and most preferably 12, 13, 14, 15, 16, 17, 18, 19 or 20 to 40 bases in length. Oligonucleotides are usually single-stranded, e.g. as probes, although the oligonucleotides may also exist in double-stranded form, e.g. used for gene mutagenesis. The oligonucleotides of the invention can be both sense (i.e. non-coding) and antisense (i.e. coding).

The term "naturally occurring nucleotides" as used in this application includes deoxyribonucleotides and ribonucleotides. The term "modified nucleotides" as used in this application includes nucleotides with a modified or substituted sugar group and the like. The term "oligonucleotide linkages" as used in this application includes such oligonucleotide linkages as phosphorothioate, dithiophosphate, selenophosphate, diselenophosphate, thioanilphosphate, anilphosphate, phosphoramidite and the like. See, e.g., LaPlanche et al. Nucci. Acids Res. 14: 9081 (1986); Stec et al. J. Am. Chem. Soc. 106: 6077 (1984); Stein et al., Nuci. Acids Res. 16: 3209 (1988); Zon et al., Anti-Cancer Drug Design 6: 539 (1991); Zon et al. Oligonucleotides and Analogues: A Practical Approach, pp. 87-108 (F. Eckstein, Oxford University Press, Oxford England (1991) ed.); Stec et al. US Patent No. 5,151,510; Uhlmann and Peyman Chemical Reviews 90: 543 (1990). The oligonucleotide may be labeled for detection, if desired.

The term "selectively hybridizes" as used herein refers to detectable and specific binding. As used herein, polynucleotides, oligonucleotides and fragments thereof selectively hybridize to nucleic acid chains under hybridization and washing conditions that minimize detectable binding to non-specific nucleic acids. Stringent hybridization conditions known in the art and discussed in this application can be used to achieve selective hybridization. Overall, sequence homology between polynes

The PL 214 003 Β1 nucleotides, oligonucleotides and fragments thereof and the selected nucleic acid sequence must be at least 80%, and usually, preferably with increasing homology, of at least 85%, 90%, 95%, 99% and 100%. Two amino acid sequences are homologous when their sequences are partially or completely identical. For example, 85% homology means that 85% of the amino acids are identical with the best alignment of the two sequences. Gaps (in one or both of the sequences being compared) are allowed to maximize matching; gaps of 5 or less are preferred, with gaps of 2 or less being more preferred. Alternatively and more preferably, the two protein sequences (or polypeptide sequences derived therefrom of at least 30 amino acids in length) are homologous, as used herein, when an alignment result obtained in the ALIGN program with a mutation data matrix and a gap penalty of 6 or more. is at least 5 (in units of standard deviation). See Dayhoff MO, in Atlas of Protein Sequence and Structure, pp. 101-110 (vol. 5, National Biomedical Research Foundation (1972)) and the second appendix to this volume, pp. 1-10. Even more preferably, two sequences or parts thereof are homologous when their amino acids are 50% or more identical when optimized by the ALIGN program.

As used herein, the term "corresponds to" means that the polynucleotide sequence is homologous (ie, identical, and not necessarily evolutionarily related) to some or all of a reference polynucleotide sequence (ie, a reference sequence), or refers to a polypeptide sequence identical to a reference polypeptide sequence. By contrast, as used herein, the term "complementary" refers to a sequence whose complementary sequence is homologous to all or part of a reference polynucleotide sequence. For example, the nucleotide sequence "TATAC" corresponds to the reference sequence "TATAC" and is complementary to the reference sequence "GTATA".

The following terms are used to describe a relationship between two or more polynucleotide or amino acid sequences: "reference sequence," comparison window, "sequence identity," percent sequence identity ", and" substantial identity. " "Reference sequence" is defined as the sequence underlying the sequence comparison; the reference sequence may be a subset of a larger sequence, for example a portion of the full length cDNA or gene sequence given in the sequence listing, or may include the complete cDNA or gene sequence. Generally, a reference sequence is at least 18 nucleotides or 6 amino acids in length, usually at least 24 nucleotides or 6 amino acids in length, and often at least 48 nucleotides or 16 amino acids in length. Since for two polynucleotides or amino acid sequences, each may (1) contain a sequence (i.e., part of a complete polynucleotide or amino acid sequence) similar in the two molecules, and (2) may also contain a sequence different in the two polynucleotides or amino acid sequences, compare the sequence two or more molecules are usually run in a "comparison window" to find and compare local regions of sequence similarity.

The term "comparison window" as used herein refers to a conceptual segment of at least 18 nucleotide or 6 amino acid consecutive positions when comparing polynucleotide or amino acid sequences to reference sequences of at least 18 nucleotides or 6 amino acids consecutive, whereby a portion of the polynucleotide sequence in the comparison window may include deletions, insertions, substitutions and the like (e.g. gaps) of 20% or less compared to the query sequence (containing no insertion or deletion) for optimal alignment of the two sequences. Optimal sequence alignment in the comparison window can be performed according to the local homology algorithm of Smith and Waterman, Adv. Appl. Math 2: 482 (1981), according to the homology alignment algorithm of Needleman and Wunsch J. Mol. Biol. 48: 443 (1970), Pearson's and Lipman's similarity method Proc. Natl. Acad. Sci (USA) 85: 2444 (1988), by computer implementations of these algorithms (software package containing GAP, BESTFIT, FASTA and TFASTA from Wisconsin Genetics Software version 7.0 (Genetics Computer Group, 575 Science Dr., Madison, Wisconsin), software package Geneworks or MacVector), or by checking and the best match (i.e., giving the highest degree of homology in the comparison pane) obtained by different methods is selected.

The term "sequence identity" as used herein means two polynucleotides or identical amino acid sequences (e.g., nucleotide by nucleotide or amino acid by amino acid) in the comparison window. As used herein, the term "percent sequence identity" is calculated for the comparison of two optimally aligned (aligned) sequences in the comparison window, by determining the number of positions having identical acid bases in both sequences.

PL 214 003 Β1 nucleic acid (e.g., A, T, C, G, U, or I) or amino acid residues to obtain the number of matching positions, dividing the number of matching positions by the number of total positions in the comparison window (i.e., by window size) and then multiplied by 100 to obtain percent sequence identity. The term "substantial identity" as used herein means polynucleotide or amino acid sequences containing sequences with at least 85% sequence identity, preferably at least 90 to 95% sequence identity, usually at least 99% identity to a reference sequence in a comparison window of at least 18 nucleotide positions (6 amino acids), often in a window of at least 24-48 nucleotide positions (8-16 amino acids), wherein the percent sequence identity is calculated by comparing the reference sequence to that sequence, which may contain deletions or insertions that number 20% or less in the reference sequence in the comparison window. The reference sequence may be a subset of a larger sequence.

The twenty common amino acid names and their abbreviations used herein follow conventional usage. See Immunology - A synthesis (2nd edition, ES Golub DR Green, eds., Sinauer Associates, Sunderland, Mass (1991)). Suitable components of the polypeptides of the invention may also be stereoisomers (e.g. amino acids D) twenty common amino acids, non-natural amino acids such as α, α-disubstituted amino acids, N-alkyl amino acids, lactic acid and other unconventional amino acids. Examples of unconventional amino acids include: 4-hydroxyproline, y-carboxyglutamate, ε-Ν, Ν, Ν-trimethylolysine, ε-Ν-acetyl lysine, O-phosphoserine, N-acetylserine, N-formylmethionine, 3-methylhistidine, 5-hydroxylysine -Ν-methyloraginine and other similar amino and imino acids (e.g. 4-hydroxyproline). In the notation of amino acids used herein, the left side corresponds to the amino terminus and the right side corresponds to the carboxyl terminus in accordance with standard notation and custom.

Similarly, unless otherwise noted, the left-hand end of a single-stranded polynucleotide sequence corresponds to the 5 'end; the left end of the double-stranded polynucleotide sequence corresponds to the 5 'end. The 5'-3 'direction of the nascent RNA transcript is referred to as the transcription direction; DNA strand sequence regions with the same sequence as the RNA strand and located 5 'to the 5' end of the transcript are referred to as "upstream sequences". upstream); DNA strand sequence regions with the same sequence as the RNA strand and located 3 'to the 3' end of the transcript are referred to as "downstream sequences."

The term "substantial identity" as used herein with respect to polypeptides means that the two polypeptide sequences, when optimally aligned, for example by GAP or BESTFIT with the default gap weights, have at least 80% sequence identity, more preferably at least 90% identity. sequence identity, more preferably at least 95% sequence identity, and most preferably at least 99% sequence identity. Preferably, amino acid residue positions that are not identical differ by conservative amino acid substitutions. Conservative amino acid substitutions refer to the replacement of residues with similar side chains. For example, a group of amino acids with aliphatic side chains is glycine, alanine, valine, lecucine, and isoleucine; a group of amino acids with aliphatic-hydroxyl side chains is serine and threonine; a group of amino acids having an amide group in their side chains is aspartate and glutamate; a group of amino acids with aromatic side chains is phenylalanine, tyrosine, andryptophan; a group of amino acids with basic side chains is lysine, arginine, and histidine; and a group of amino acids with sulfur-containing side chains is methionine and cysteine. The valine-leucine-isoleucine, phenylalanine-tyrosine, lysine-arginine, alanine-valine, aspartate-glutamate, and glutamine-asparagine groups are preferred conservative amino acid substitution groups.

As discussed herein, minor changes in the amino acid sequence of the antibodies or immunoglobulin molecules are contemplated to be within the scope of the present invention such that amino acid sequence differences are kept to at least 90%, 95%, and most preferably 99%. In particular, conservative amino acid substitutions are contemplated. Conservative amino acid substitutions are those that take place within a family of side chain related amino acids. The genetically coded amino acids are generally classified into the families (1) acidic = aspartate, glutamate; (2) basic = lysine, arginine, histidine; (3) nonpolar = alanine, valine, leucine, isoleucine, proline, phenylalanine, methionine, tryptophan and (4) uncharged polar = glycine, asparagine, glutamine, cysteine, serine, threonine and tyrosine. The most advantageous

PL 214 003 Β1 families are: serine and threonine, which are the aliphatic-hydroxyl amino acid family; asparagine and glutamine which are a family of amino acids containing an amide group; alanine, valine, leucine, and isoleucine, which are an aliphatic family; and phenylalanine, tryptophan, and tyrosine, which constitute the family of aromatic amino acids. For example, a single replacement of leucine with isoleucine or valine, aspartate with glutamate, threonine with serine, or a similar replacement of an amino acid with a structurally related amino acid may be expected to have little effect on the binding properties of the resultant molecule, particularly if the substitution does not involve the amino acid at the binding site. By determining the specific activity of a polypeptide derivative, it can be readily determined whether a functional polypeptide is formed as a result of an amino acid change. Suitable tests are described in detail in this application. One skilled in the art can readily prepare fragments or analogs of antibodies or immunoglobulin molecules. Preferably, the amino and carboxyl termini of the fragments or analogs are close to the boundaries of the functional domains. Structural and functional domains can be determined by comparing nucleotide and / or amino acid sequence data with public or private databases. Preferably, computerized comparative methods are used to determine sequence motifs or predicted protein conformation domains found in other proteins of known structure and / or function. Methods for identifying protein sequences that fold into known three-dimensional structures are known. Bowie et al., Science 253: 164 (1991). Thus, the following examples show that one skilled in the art can recognize sequence and structural conformation motifs that can be used to define structural and functional domains.

Preferred amino acid substitutions are those that: (1) reduce the susceptibility to proteolysis, (2) reduce the oxidability, (3) change the binding affinity for the formation of protein complexes, and (4) confer or alter other physico-chemical or functional properties of such analogs. Analogs can include various muteins of the sequence other than naturally occurring peptide sequences. For example, single or multiple amino acid substitutions (preferably conservative amino acid substitutions) may be made in a naturally occurring sequence (preferably in a portion of the polypeptide outside of the domain (s) involved in inter-molecule interactions). A conservative amino acid substitution should not substantially alter the structural properties of the parent molecule (e.g. the amino acid change should not break the helix found in the parent molecule or disrupt other types of secondary structures found in the parent molecule.) Examples of art-known secondary and tertiary protein structures are described in Proteins, Structures and Molecular Principles (Creighton, WH Freeeman and Company, New York (1984)); Introduction to Protein Structure (C. Branden and J. Tooze, Garland Publishing, New York, NY (1991)) and Thornton et al., Nature 354: 105 (1991).

The term "polypeptide fragment" as used herein refers to a polypeptide that is deleted at the amino acid and / or carboxy terminus, but wherein the sequence of the remaining amino acids is identical to the corresponding positions of a naturally occurring sequence determined, for example, by the full length cDNA sequence. Fragments are usually at least 5, 6, 8 or 10 amino acids in length, preferably at least 14 amino acids in length, more preferably at least 20 amino acids in length, typically at least 50 amino acids in length, and even more preferably at least 70 amino acids in length. The term "analog" as used herein refers to polypeptides consisting of a stretch of at least 25 amino acids with substantial identity to part of the deduced amino acid sequence and with at least one of the following properties: (1) specific binding to CTLA-4, under appropriate binding conditions, (2) blocking ability binding of CTLA-4 to its receptors, or (3) the ability to inhibit growth of cells expressing CTLA-4 in vitro or in vivo. Typically, polypeptide analogs contain an amino acid substitution (or addition or deletion) conserved with respect to the naturally occurring sequence. Analogs are typically 20 amino acids in length, preferably at least 50 amino acids or longer, and can often be as long as a naturally occurring polypeptide.

Peptide analogs are often used in the pharmaceutical industry as non-peptide drugs with properties analogous to the peptide matrix thereof. This type of non-peptide compound is called "peptide mime ethics" or "peptidomimetics". Fauchere, J. Adv. Drug Res. 15:29 (1986); Vebere and Freidinger TINS ​​p. 392 (1985); Evans et al. J. Med. Chem. 30: 1229 (1987). Such compounds are typically developed using computer molecular modeling. Peptide mimetics that are structurally similar to therapeutically useful peptides may be used

PL 214 003 Β1 is important to produce an equivalent prophylactic or therapeutic effect. In general, peptidomimetics are structurally similar to a model peptide (e.g., a polypeptide with biochemical properties or pharmacological activity), such as a human antibody, but one or more of their peptide bonds are optionally replaced with a bond selected from -CH₂NH-, -CH₂S-, -CH₂-CH₂-, -CH = CH- (cis and trans), -COCH₂-, -CH (OH) CH₂- and -CH₂SO-, by methods well known in the art. The ordered substitution of one or more amino acids of a consensus sequence (consensus sequence) with a D-amino acid of the same type (e.g., substituting D-lysine for L-lysine) can be used to produce a more stable peptide. In addition, internally linked peptides containing a consensus sequence (Rizo and Gierasch Ann Rev. Biochem. 61: 387 (1997)); for example, by adding internal cistern residues that allow the formation of disulfide bridges that lead to protein cyclization.

"Antibody or" antibody peptide (s) "refers to an intact antibody or a binding fragment thereof (ie, a binding fragment) that competes with the intact antibody for specific binding. Binding fragments are produced by recombinant DNA techniques or by chemical or enzymatic cleavage of intact antibodies. Binding fragments include Fab, Fab ', F (ab')₂, Fv, and single chain antibodies. By an antibody other than "bispecific or" bifunctional, it is meant an antibody for which each binding site is identical. An antibody substantially inhibits receptor-ligand binding when an excess of the antibody reduces the amount of ligand-bound receptor by at least 20%, 40%, 60%, 80%, and usually more than 85% or more (measured in injection in a competitive binding assay).

The term "epitope" includes any protein determinant capable of specifically binding to an immunoglobulin or T-cell receptor. Epitope determinants typically consist of chemically active surface groupings of molecules such as amino acids or sugar side chains, and typically possess specific three-dimensional properties as well as specific charge distribution. . An antibody is considered to specifically bind an antigen if the dissociation constant is <1 µΜ, preferably <100 nM, and most preferably <10 nM.

The term "agent" as used herein refers to a chemical compound, a mixture of chemicals, a biological macromolecule, or an extract made from biological material.

As used herein, the term "tag" or "labeled" refers to the inclusion of a detectable marker, e.g., by inclusion of a radiolabeled amino acid, or the addition of biotinyl moieties to the polypeptide that can be detected with labeled avidin (e.g. by optical or colorimetric methods). In some situations, the marker or marker can also be therapeutic. A variety of methods for labeling polypeptides and glycoproteins known in the art can be used. Examples of tags for polypeptides include, but are not limited to, the following: radioisotopes or radionuclides (e.g.³H, ¹⁴C, ¹⁵N, ³⁵S, ⁹⁰Y, Tc, 'in, ¹²⁵l, ¹³¹1), fluorescent markers (e.g. FITC, rhodamine, lanthanide doped phosphors), enzymatic labels (e.g. horseradish peroxidase, β-galactosidase, luciferase, alkaline phosphatase), chemiluminescent, biotinyl groups, pre-defined polypeptide epitopes recognized by a secondary reporter group (e.g., leucine zipper sequences, binding sites for secondary antibodies, metal binding domains, epitope tags). The labels may be attached by spacer arms of various lengths to reduce potential spherical hindrance.

The term "pharmaceutical agent or medicament" as used herein refers to a chemical compound or composition capable of producing a desired therapeutic effect when properly administered to a patient. Other chemical terms are used herein according to conventional usage and their meaning in the art as indicated, for example, in The MaeGraw-Hill Dictionary of Chemical Terms (Parker S., McGraw-Hill, San Francisco (1985)).

The term "anti-neoplastic agent" as used herein refers to agents whose functional property is to inhibit the development or progression of a human tumor, in particular malignant neoplastic (cancerous) lesions such as cancer, sarcoma, lymphoma or leukemia. Inhibition of metastasis is a common property of anti-cancer agents.

As used herein, the term "substantially pure" means that the type of substance is predominantly present (e.g., its molar mass is greater than any of the other individual components of the composition), and preferably the substantially pure fraction is a composition in which the type of substance is at least 50%. (on a molar basis) all present are born

PL 214 003 Β1 of macromolecules. Typically a substantially pure composition comprises greater than about 80% of all types of macromolecules present in the composition, more preferably greater than about 80%, 90%, 95%, and 99%. Most preferably, the substance of interest is purified to substantial homogeneity (impurities cannot be detected in the mixture by conventional detection methods) and the composition consists essentially of one type of macromolecule.

The term patients includes humans and veterinary actors.

Structure of antibodies

As is known, the basic structural unit of an antibody is a tetramer. Each tetramer consists of two identical pairs of polypeptide chains, each pair of a "light chain (approximately 25 kDa) and a" heavy chain (approximately 50-70 kDa). The amino-terminal portion of each chain contains a variable region of 100 to 110 or more amino acids primarily responsible for antigen recognition. The carboxy-terminal portion of each chain is the constant region primarily responsible for effector functions. Human light chains are classified as kappa and lambda chains. Heavy chains are classified as mu, delta, gamma, alpha, or epsilon, and define the antibody's isotype as IgM, IgD, IgG, IgA, and IgE, respectively. Within the light and heavy chains, the variable regions are joined by a "J" region of about 12 or more amino acids, and the heavy chain also includes a "D" region of about 10 amino acids. See Fundamental Immunology Chap. 7 (Paul, W., 2nd ed. Raven Press, NY (1989)). The variable regions from each light / heavy chain pair constitute the antibody combining site.

Thus, an intact IgG antibody has two binding sites. Except for bifunctional or bispecific antibodies, the two binding sites are the same.

All chains share the same general structure of relatively conserved framework regions (FRs) joined by three hypervariable regions, also called complementarity determining regions or CDRs. complementarity determining regions). The CDRs of the two chains of each pair are aligned with each other through the framework regions, allowing the binding of a specific epitope. In the N-terminus to C-terminus, both the light and heavy chains contain the FR1, CDR1, FR2, CDR2, FR3, CDR3, and FR4 domains. Amino acids are assigned to the appropriate domain as defined by Kabat Sequences of Proteins of Immunological Interest (National Institutes of Health, Bethesda, Md. (1987 and 1991)) or Chothia and Lesk J., Mol. Biol. 196: 901-917 (1987); Chothia et al. Nature 342: 878-883 (1989).

Bispecific or bifunctional antibodies are artificial hybrid antibodies having two different heavy / light chain pairs and two different binding sites. Bispecific antibodies can be made by a variety of methods, including fusion of hybridomas or fusion of Fab 'fragments. See, e.g., Songsivilai and Lachmann, Clin. Exp. Immunol. 79: 315-321 (1990), Kostelny et al., J. Immunol. 148: 1547-1553 (1992). In addition, bispecific antibodies can be produced as "diabodies". diabody) (Holliger et al. "Diabodies: smali bivalent and bispecific antibody fragments" PNAS USA 90: 6444-6448 (1993)) or "Janusin" type antibody (Traunecker et al. "Bispecific single chain molecules (Janusins) target cytotoxic lymphocytes on HIV infected cells "EMBO J 10: 3655-3659 (1991) and Traunecker et al." Janusin: new molecular design for bispecific reagents "Int J CancerSuppl 7: 51-52 (1992)).

The production of bispecific antibodies can be a relatively laborious process compared to the production of conventional antibodies, and the yield and degree of purity are generally lower for bispecific antibodies. Bispecific antibodies do not exist as fragments having a single binding site (e.g., Fab, Fab ', and Fv).

Human antibodies and the humanization of antibodies

Human antibodies avoid some of the problems associated with antibodies having murine and rat variable and / or constant regions. The presence of such mouse and rat derived proteins may lead to the rapid clearing of antibodies from the body or may lead to an immune response in the patient against the antibody. In order to avoid the use of mouse or rat derived antibodies, it has been postulated that it is possible to obtain humanized antibodies or to generate fully human antibodies by introducing human antibody functions into rodents so that the rodent can produce antibodies having fully human sequences.

PL 214 003 Β1

Human antibodies

The ability to clone and reconstruct human loci millions of bp in the YAC and introduce them into mouse germlines is a powerful tool for identifying functional components of very large or only pre-mapped loci, as well as for generating useful models of human disease. Furthermore, using such technology to replace mouse loci with human equivalents could provide a unique opportunity to study expression and regulation of gene products during development, their communication with other systems, and their contribution to disease induction and development.

An important practical application of such a strategy is to "humanize" the mouse humoral immune system. Introducing human immunoglobulin (Ig) loci into mice in which endogenous Ig genes have been inactivated offers an opportunity to study the mechanisms underlying the programmed expression and assembly process of antibodies, as well as their role in B cell development. Moreover, such a strategy would be an ideal source for the production of fully human monoclonal antibodies (Mabs) - a quantum leap towards the promise of antibody therapy of human disease. Fully human antibodies are expected to minimize the immunogenic and allergic responses associated with murine and mouse Mabs and thus increase the efficacy and safety of the administered antibodies. The use of fully human antibodies can be expected to provide significant benefits in the treatment of chronic and relapsing human diseases such as inflammation, autoimmune diseases and cancer, which require repeated administration of antibodies.

One approach to this goal has been to obtain mice that do not produce murine antibodies (i.e. deficient in self-antibody production) with large fragments of human Ig loci, expecting such mice to produce a wide repertoire of human antibodies in the absence of murine antibodies. Large human Ig fragments retain high gene variability as well as appropriate regulation of antibody production and expression. By using the mouse machinery to obtain diversity and selection of antibodies, and the intolerance to human proteins, the reconstituted human antibody repertoire in these mice can yield high affinity antibodies to any antigen of interest, including human antigens. Human Mabs having the desired antigen specificity can be generated and selected on the spot using hybridoma technology.

This overall strategy was presented in connection with our development of the first type of XenoMouse ™ as published in 1994. See Green et al. Nature Genetics 7: 13-21 (1994). XenoMouse ™ obtained with the use of yeast artificial chromosomes ( yeast artificial chromosomes (YAC) containing 245 kb and 190 kb fragments for the germline configuration of the human heavy chain and kappa light chain, respectively, which contain variable and constant region core sequences. Human Ig containing YAC has been shown to correspond to the mouse system in both rearrangement and expression of antibodies, and has the ability to replace inactivated murine genes. This has been demonstrated by their ability to induce B cell development, to generate a repertoire of adult all-human antibodies, and to produce antigen-specific human Mabs. These results also suggest that the introduction of more human Ig loci containing more V genes, additional regulatory elements and human Ig constant regions can recreate the substantially complete repertoire that is characteristic of the human humoral response to infection and immunization. The work of Green et al. has recently been expanded to incorporate over 80% of the human antibody repertoire by introducing thousands of bp YAC fragments and the germline configuration of the human heavy chain and kappa light chain loci, respectively, to generate the XenoMouse ™ mice. See Mendez et al. Nature Genetics 15: 146-156 (1997), Green and Jakobovits J: Exp. Med. 188: 483-495 (1998) and U.S. Patent Application Serial No. 08 / 759,620, filed December 3, 1996.

This approach is further discussed and set forth in U.S. Patent Applications Serial Numbers: 07 / 466,008, January 12, 1990, 07 / 610,515, November 8, 1990, 07 / 919,297, July 24, 1992, 07 / 922,649, July 30, 1992, 08 / 031,801, March 15, 1993, 08 / 112,848 of August 27 1993, 08 / 234,145 of 28 April 1994, 08 / 376,279, of 20 January 1995, 08/430, 938 of 27 April 1995, 08 / 464,584 of 5 June 1995, 08 / 464,582 of 5 June 1995, 08 / 463,191 of 5 June 1995, 08 / 462,837 of 5 June 1995, 08 / 486,853 of 5 June 1995, 08 / 486,857 of 5 June 1995, 08 / 486,859 of 5 June 1995, 08 / 462,513 filed June 5, 1995, 08 / 724,752 filed October 2, 1996, and 08 / 759,620 filed December 3, 1996. See also Mendez et al. Nature Genetics 15: 146-156 (1997) and Green and Jakobovits J. Exp. Med. 188: 483495

PL 214 003 Β1 (1998). See also European Patent Application No. EP 0463 151 B1, issued June 12, 1996, International Patent Application No. WO 94/02602, published February 3, 1994, International Patent Application No. WO 96/34096, published October 31, 1996 and WO 98 / 24,893 published June 11, 1998.

In an alternative approach, others, including GenPharm International, Inc. they used the "minilocus" strategy. In the minilocus strategy, an exogenous Ig locus is mimicked by the inclusion of pieces (individual genes) derived from the Ig locus. Thus, one or more genes of V_H.one or more genes for D_H. one or more J genes_H., the mu constant region and the second constant region (preferably the gamma constant region) are created in a construct for insertion into an animal. This approach is described in U.S. Patent No. 5,545,807 to Surani et al. and U.S. Patent Nos. 5,545,806, 5,625,825, 5,625,126, 5,633,425, 5,661,016, 5,770,429, 5,789,650, and 5,814,318 each to Lonberg and Kay, U.S. Patent No. 5,591,669 to Krimpenfort and Bems, U.S. Patent Nos. 5,612,315675 to 5,789,215675, Bems et al., In U.S. Patent No. 5,643,763 to Choi and Dunn, and in U.S. Patent Applications, GenPharm International, serial numbers: 07 / 574,748 of August 29, 1990, 07 / 575,962 of August 31, 1990, 07 / 810,279 of December 17, 1991, 07 / 853,408 of March 18, 1992, 07 / 904,068, filed June 23, 1992, 07 / 990,860, filed December 16, 1992, 08 / 053,131 of 26 April 1993, 08 / 096,762 of 22 July 1993, 08 / 155,301 of 18 November 1993, 08 / 161,739 of 3 December 1993, 08 / 165,699 of 10 December 1993, 08 / 209,741 of 9 March 1994. See also European Patent Application No. 0 546 073 B1, International Patent Application No. WO 92/03918, WO 92/22645, WO 92/22647, WO 92/22670, WO 93/12227, WO 94/00569, WO 94/25585, WO 96/14436, WO 97/13852 and WO 98/24884. See also Taylor et al. (1992), Chen et al. (1993), Tuaillon et al., 1993, Choi et al. (1993), Lonberg et al. (1994), Taylor et al. (1994) and Tuaillon et al. (1995), Fishwild et al. (1996).

Surani et al., Who transferred rights from the invention to the Medical Research Counsel ("MRC"), generated a transgenic mouse bearing the Ig locus by using the minilocus approach. Lonberg and Kay of GenPhann International, following in the footsteps of the present inventors, proposed inactivating the endogenous mouse Ig locus coupled with essentially the work of Surani et al.

The advantage of the minilocus approach is the speed with which constructs containing portions of the Ig locus can be produced and introduced into animals. However, on the other hand, a significant disadvantage of the minilocus approach is that in theory a small number of V, D and J genes are insufficiently differentiated. Indeed, the published papers seem to support this concern. B-cell recovery and antibody production by minilocus animals appeared to be inhibited. Therefore, research focussed on the present invention has been steadily directed towards introducing larger portions of the Ig locus in order to achieve greater diversity and to reconstruct the immune repertoire of animals.

Human anti-mouse antibody (HAMA) responses have led to the industrial production of chimeric and otherwise humanized antibodies. Although chimeric antibodies have human constant regions and murine variable regions, it would be expected that some human anti-chimeric antibody (HACA) responses would be observed, especially with long-term or multi-dose administration of the antibody. Thus, it will be desirable to provide fully human antibodies to CTLA-4 to alleviate the concerns and / or effects of a HAMA or HACA response.

Technologies of humanization and presentation

As discussed above in relation to human antibody production, there are advantages to producing antibodies with reduced immunogenicity. To some extent, they can be obtained in conjunction with humanization and display techniques using appropriate libraries. It will be appreciated that murine antibodies or antibodies from other species can be humanized or primatized using techniques known in the art. See, for example, Winter and Harris Immunol Today 14: 43-46 (1993) and Wright et al. Crit. Reviews in Immunol 12125-168 (1992). The antibody of interest can be constructed by recombinant DNA technology to replace the CH1, CH2, and CH3 hinge domains and / or the framework domains with the corresponding human sequences (see WO 92/02190 and US Patent Nos. 5,530,101, 5,585,089, 5,693,761, 5,693,792, 5,714,350 and 5,777,085). Furthermore, it is known in the art to use Ig cDNA to reconstruct immunoglobulin genes (Liu et al. PNAS 84: 3439 (1987) and J. Immunol. 139: 3521 (1987)). The mRNA is isolated from the hybridoma or other antibody-producing cells and used to generate the cDNA. cDNA being the subject of in

PL 214 003 Β1 teering can be amplified in the polymerase chain reaction using specific primers (US Patent Nos. 4,683,195 and 4,683,202). Alternatively, libraries are generated and screened to isolate a sequence of interest. The DNA sequence encoding the antibody variable region is fused to the human constant region sequences. Human constant region gene sequences can be found in Kabat et al. (1991) Sequences of Proteins of Immunological Interest, NIH Publication No. 91-3242. Human C region genes are readily available from known clones. The selection of an isotype will be guided by desired effector functions such as complement fixation or activity in antibody dependent cellular cytotoxicity. The preferred isotypes are IgG1, IgG2, IgG3 and IgG4. IgG2 and IgG4 are particularly useful isotypes for the antibodies described herein. Any of the human light chain, kappa or lambda constant regions can be used. The chimeric humanized antibodies can then be expressed using conventional methods.

Antibody fragments such as Fv, F (ab ')₂ and Fabs can be prepared by cleaving the protein of interest, e.g., by protease cleavage or chemical cleavage. Alternatively, a truncated gene is designed. For example, a chimeric gene encoding part of an F (ab ') fragment₂it will contain DNA sequences encoding the CHI domain and the H chain hinge region followed by a translation stop codon so as to obtain a truncated molecule.

One approach may use consensus sequences encoding the heavy and light chains of the J regions to design oligonucleotides for use as primers to insert useful restriction sites into the J region for the subsequent connection of V region fragments with human C region stretches. modified by site-directed mutagenesis to place the restriction site in an analogous position in the human sequence.

Expression vectors include plasmids, retroviruses, cosmids, YACs, EBV-derived episomes, and the like. A convenient vector is one that encodes a functionally complete human immunoglobulin CH or CL sequence, with appropriate restriction sites engineered so that any VH or VL sequence may be inserted and expressed. In such vectors, the splice typically occurs between the splice donor site in the inserted J region and the splice acceptor site that precedes the human C region, and in the splice region that occurs within human CH exons. Polyadenylation and transcription termination occur at native chromosomal sites downstream of the coding regions. The resulting chimeric antibody can be attached to any strong promoter, including retroviral LTR such as, e.g., the SV-40 early promoter (Okayama et al. Mol. Cell. Bio. 3: 280 (1983)), Rous sarcoma virus LTR ( Gorman et al. PNAS 79: 6777 (1982) and Moloney murine leukemia virus LTR (Grosschedl et al. Cell 41: 885 (1985)); native Ig promoters, etc.

In addition, human antibodies or antibodies from other species can be produced by display technology, including but not limited to phage display, retrovirus display, ribosome display, and other techniques used in the art, and the resulting molecules can be subjected to further maturation. such as affinity maturation as these techniques are well known in the art. Wright and Harris, supra., Hanes and Plucthau PNAS USA 94: 4937-4942 (1997) (ribosome display), Parmley and Smith, Gene 73: 305-318 (1988) (phage display), Scott, TIBS 17 : 241-245 (1992), Cwirla et al., PNAS USA 87: 6378-6382 (1990), Russel et al. Nuci. Acids Research 21: 1081-1085 (1993), Hoganboom et al. Immunol. Reviews 130: 43-68 (1992), Chiswell and McCafferty TIBTECH 10: 80-84 (1992) and US Patent No. 5,733,743. If display technology is used to generate non-human antibodies, such antibodies can be humanized as described above.

Using these techniques, antibodies can be generated against CTLA-4 expressing cells, CTLA-4 itself, CTLA-4 forms, peptides and epitopes thereof, and expression libraries (see, e.g., US Patent No. 5,703,057), which can then be screened as described above for those described above. above activity.

Additional Criteria for Antibody Drugs

As will be appreciated, it is generally undesirable to kill CTLA-4 expressing cells. Rather, it is generally desirable to simply inhibit CTLA-4 binding to its ligands in order to attenuate the down-regulation of T cells. One of the major mechanisms by which antibodies kill cells is through complement fixation and participation in CDC. The constant region of the antibody plays an important role in the ability of the antibody to fix complement and participate in CDC. So gene

Orally, the isotype of the antibody may be selected to provide complement fixation capacity or not. For the present invention, generally, as mentioned above, it is not preferred to use an antibody that kills cells. There are many isotypes of antibodies that are capable of complement and CDC binding, including but not limited to: mouse IgM, mouse IgG2a, mouse IgG2b, mouse IgG3, human IgM, human IgG1, and human IgG3. Such isotypes do not include, but are not limited to, human IgG2 and human IgG4.

It will be appreciated that the antibodies that are produced need not initially have the desired isotype, but rather the antibody produced may be of any isotype that can later be switched using conventional techniques known in the art. Such techniques include, but are not limited to, the use of direct recombination techniques (see, e.g., US Patent No. 4,816,397). cell-cell fusion techniques (see, e.g., Patent Application No. 08 / 730,639 filed October 11, 1996).

In the cell-cell fusion technique, a myeloma or other cell line is prepared which has a heavy chain with the desired isotope and another myeloma cell line or another which has a light chain. Such cells can then be fused and the cell line expressing the intact antibody isolated.

As an example, the majority of anti-CTLA-4 antibodies discussed herein are human anti-CTLA-4 IgG2 antibody. Since such antibodies have the desired binding ability to CTLA-4 molecules, any such antibodies can be readily isotyped to generate, for example, the human IgG4 isotype while still having the same variable region (which defines the specificity of the antibody and some of its affinity).

Thus, once antibodies have been generated which possess the desired "structural" attributes as discussed above, they can be provided with at least some additional desirable "functional" attributes by isotype alteration.

Design and manufacture of other drugs

The design of other therapeutic agents, including other antibodies, other antagonists, or non-antibody chemical molecules is facilitated in accordance with the present invention and based on the activity of the antibodies that are generated and characterized in relation to CTLA-4. Such agents include, but are not limited to, antibodies with similar binding activity or functionality, advanced antibody-based drugs such as bispecific antibodies, immunotoxins, and radiolabeled drugs, peptide drug production, gene therapies, and in particular, intrabody production of intracellular antibodies. antisense and small molecule therapies. In addition, as discussed above, the effector functions of the antibodies of the invention can be altered by isotype change to IgG1, IgG2, IgG3, IgG4, IgD, IgA, IgE, or IgM for various therapeutic applications.

With the development of advanced therapeutics based on antibodies for which complement fixation is a desirable attribute, it is possible to shift the complement dependency aside in favor of cell killing by using, for example, bispecific antibodies, immunotoxins and radiolabeled drugs.

With respect to bispecific antibodies, bispecific antibodies can be made that include (i) two antibodies linked together: one specific for CTLA-4 and the other for another molecule, (ii) a single antibody that has one chain specific for CTLA-4 and a second chain specific for another molecule; or (iii) a single chain antibody that has specificity for CTLA-4 and another molecule. Such bispecific antibodies can be made using well known techniques, for example for (i) and (ii), see Fanger et al. Immunol Methods 4: 72-81 (1994) and Wright and Harris, supra, and for (iii) see Traunecker et al. Int. J. Cancer (Suppl.) 7: 51-52 (1992).

In addition, "kappabodies) can also be produced (III et al." Design and construction of a hybrid immunoglobulin domain with properties of both heavy and light chain variable regions "Protein Eng 10: 949-57 (1997)) , "Mini-bodies" (Martin et al. "The affinity-selection of a minibody polypeptide inhibitor of human interleukin-6" EMBO J 13: 5303-9 (1994)), "di-bodies" ( Holliger et al. ,, 'Diabodies': "smali bivalent and bispecific antibody fragments" PNAS USA 90: 6444-6448 (1993)), or antibodies of the type "Janusin" (Traunecker et al. "Bispecific single chain molecules (Janusins) target cytotoxic lymphocytes on HIV infected cells "EMBO J 10: 3655-3659 (1991) and Traunecker et al." Janusin: new molecular design for bispecific reagents "Int J Cancer Suppl 7: 51-52 (1992)).

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With regard to immunotoxins, antibodies can be modified to act as immunotoxins using techniques well known in the art. See, for example, Vitetta Immunol Today 14: 252 (1993). See also US Patent No. 5,194,594. For the production of radiolabeled antibodies, such modified antibodies can be readily obtained using techniques well known in the art. See, e.g., Junghans et al. in Cancer Chemotherapy and Biotherapy 655-686 (2nd ed., Chafner and Longo, eds., Lippincott Raven (1996)). See also U.S. Patent Nos. 4,681,581, 4,735,210, 5,101,827, 5,102,990 (RE 35,500), 5,648,471, and 5,697,902. Any of the immunotoxins and radiolabeled molecules will likely kill CTLA-4 expressing cells, and particularly those cells for which the antibodies described herein are effective.

With respect to the production of peptide drugs, by using structural information related to CTLA-4 and antibodies directed against it, such as the antibodies of the invention (as discussed below with respect to small molecules) or by screening peptide libraries, anti-CTLA-therapeutic peptides can be produced. 4. Design and screening of peptide libraries is discussed in Houghten et al. Biotechniques 13: 412-421 (1992), Houghten PNAS USA 82: 5131-5135 (1985), Pinalla et al. Biotechniques 13: 901-905 (1992), Blake and Litzi-Davis BioConjugate Chem. 3: 510-513 (1992). Immunotoxins and radiolabeled drugs can also be produced, in a similar manner, in combination with peptide moieties, as discussed above for antibodies.

Important information regarding the binding of an antibody to an antigen can be obtained through phage display experiments. Such experiments are generally performed by screening a phage library expressing random peptides for binding of antibodies of the invention to determine whether binding peptides can be isolated. If successful, some epitope information can be gleaned from the binding peptides.

In general, phage libraries expressing random peptides based on the M13 bacteriophage system can be purchased from New England Biolabs (7-mer and 12-mer libraries, Ph.D.-7 Peptide 7-mer Library Kit and Ph.D.-12 Peptide 12 -mer Library Kit, respectively). The 7-mer library contains 2.0 × 10⁹ independent clones that represent most, if not all, of the 20⁷ = 1.28 x 10⁹ possible 7mer sequences. A 12-mer library is approximately 1.9 × 10⁹ independent clones, representing only a small sample of the possible 20¹² = 4.1 χ 10¹⁵ 12-mer sequences. Each of the 7-mer and 12-mer libraries are screened or screened according to the manufacturer's instructions and the plates are coated with antibodies to capture the appropriate antibody (e.g., goat anti-human IgG Fc for an IgG antibody) and then washed. Bound phages are eluted with 0.2 M glycine-HCl, pH 2.2. After 3 rounds of selection / expansion at constant stringency (0.5% Tween), clones from the libraries that react with one or more antibodies can be characterized by DNA sequencing. The reactivity of the peptides can be confirmed by ELISA. Additional discussion of epitope analysis in peptides can be found in Scott, JK and Smith, GP Science 249: 386-390 (1990); Cwirla et al. PNAS USA 87: 63786382 (1990); Felici et al. J. Mol. Biol. 222: 301-310 (1991) and Kuwabara et al. Nature Biotechnology 15: 74-78 (1997).

The present invention also facilitates the design of gene and / or antisense drugs by employing conventional techniques. Such agents can be used to modulate the function of CTLA-4. Accordingly, the antibodies of the present invention facilitate the development and use of functional assays. The design and strategy of therapies based on antisense molecules is discussed in detail in International Patent Application No. WO 94/29444. Gene therapy design and strategies are well known. However, the use of gene therapy techniques involving intrabody antibodies could prove to be particularly advantageous. See, e.g., Chen et al. Uman Gene Therapy 5: 595-601 (1994) and Marasco Gene Therapy 4:11-15 (1997). The overall picture and issues relating to gene drugs are discussed in the international patent application No. WO 97/38137. The genetic material encoding the antibody (such as 4.1.1,4.8.1 or 6.1.1 and others) may be contained in an appropriate expression system (be it viral, weakened virus based, non-viral, naked DNA, or otherwise) and administered to the host to produce antibodies in vivo in the host.

It is also possible to develop small therapeutic molecules. Based on the present invention, drugs can be designed to modulate CTLA-4 activity. The knowledge gained from the structure of the CTLA-4 molecule and its interactions with other molecules, such as the antibodies of the invention and other antibodies disclosed herein, CD28, B7, B7-1, B7-2, can be used to theoretically design additional therapeutic agents. In this aspect, for the purpose of purchase20

In drug design efforts, theoretical techniques of drug design such as X-ray crystallography, computer assisted molecular modeling (CAMM), quantitative or qualitative structure-function relationship (CAMM) can be applied. quantitative or qualitative structure-activity relationship (QSAR) and similar technologies. Theoretical design makes it possible to predict protein or synthetic structures that can interact with a molecule or specific forms thereof that can be used to modify or modulate CTLA-4 activity. Such structures can be chemically synthesized or expressed in biological systems. Such an approach is discussed in Capsey et al. Genetically Engineered Human Therapeutic Drugs (Stockton Press, NY (1988)). In fact, theoretical design of molecules (be they peptides, peptidomimetics, small molecules, or the like) based on a known or established structure-activity relationship with other molecules (such as the antibodies of the invention) becomes routine. See, e.g., Fry et al, "Specific, irreversible inactivation of the epidermal growth factor receptor and erbB2, by a new class of tyrosine kinase inhibitor" Proc Natl Acad Sci LFSA 95: 12022-7 (1998); Hoffman et al. "A model of Cdc25 phosphatase catalytic domain and Cdk-interaction surface based on the presence of a rhodanese homology domain" J Mol Biol 282: 195-208 (1998); Ginalski et al. "Modeling of active forms of protein kinases: p38-a case study" Acta Biochim Pol 44: 557-64 (1997); Jouko et al. "Identification of csk tyrosine phosphorylation sites and a tyrosine residue important for kinase domain structure" Biochem J 322: 927-35 (1997); Singh et al. "Structure-based design of a potent, selective, and irreversible inhibitor of the catalytic domain of the erbB receptor subfamily of protein tyrosine kinases" J Med Chem 40: 1130-5 (1997); Mandel et al. "ABGEN: a knowledge-based automated approach for antibody structure modeling" Nat Biotechnol 14: 323-8 (1996); Monfardini et al, "Rational design, analysis, and potential utility of GM-CSF antagonists" Proc Assoc Am Physicians 108: 420-31 (1996); Furet et al. "Modeling study of protein kinase inhibitors: binding modę of staurosporine and origin of the selectivity of CGP 52411" J Comput Aided Mol Des 9: 465-72 (1995).

In addition, combinatorial libraries can be designed and synthesized and used in screening programs, such as large-scale screening attempts.

Drug administration and pharmaceutical compositions

It will be appreciated that administration of the therapeutic agents can be accomplished by administering them together with suitable carriers, excipients, and other agents that are included in the composition to provide improved delivery, delivery, tolerance, and the like. Numerous suitable compositions can be found in a body known to those skilled in the art of pharmaceutical chemistry: Remington's Pharmaceutical Sciences (2nd ed. 15, Mack Publishing Company, Easton, PA (1975)), especially in chapter 87 by Blaug, Seymour. These compositions include, for example, powders, pastes, ointments, gels, waxes, oils, lipids, lipid containing vesicles (cationic or anionic) (such as LipofectinTM), DNA conjugates, anhydrous absorption pastes, oil-in-water or oil-water emulsions. carbon wax emulsions (polyethylene glycols of different molecular weights), semi-liquid gels, semi-liquid mixtures containing carbon waxes. Any of the above mixtures may be suitable for therapy and therapy provided that the active ingredient is not inactivated by the composition and that it is physiologically acceptable and tolerated by the route of administration. See also Powell et al. "Compendium of excipients for parenteral formulations" PDA J Pharm Sci Technol. 52: 238-311 (1998) and references cited therein for additional information regarding excipients and carriers well known in the art of drug chemistry. Production of antibodies

The antibodies of the invention are preferably prepared by using a transgenic mouse into which a substantial portion of the human genome necessary for antibody production has been introduced and at the same time rendered incapable of producing endogenous murine antibodies. Such mice are then capable of producing human immunoglobulin and antibody molecules, and are incapable of producing murine immunoglobulin and antibody molecules. The technologies used to achieve this goal are disclosed in patents, patent applications, and references in the related art section. However, particularly preferred embodiments for the process of producing transgenic mice, and antibodies therefrom, are disclosed in US Patent Application Serial No. 08 / 759,620 filed on December 3, 1996. See also Mendez et al. Nature Genetics 15: 146-156 (1997).

Thanks to the use of such technology, we have created fully human monoclonal antibodies to various antigens. Basically, we immunize XenoMouse ™ mouse lines with the antigen of interest, take lymphatic cells (such as B cells) from mice that express antibodies, connect such extracted cells with myeloid cell lines

PL 214 003 Β1 ka to obtain immortalized hybridoma cell lines, and such hybridoma cell lines are screened and selected to identify hybridoma cell lines that produce antibodies specific for the antigen of interest. We used these techniques to generate CTLA-4 specific antibodies. Herein we describe the generation of multiple hybridoma cell lines that produce CTLA-4 specific antibodies. In addition, we have provided characterization of the antibodies produced by such cell lines, including analysis of the amino acid sequence of the heavy and light chains of such antibodies.

Antibodies derived from the hybridomas discussed herein are designated 3.1.1, 4.1.1, 4.8.1,4.10.2, 4.13.1,4.14.3, 6.1.1, 11.2.1, 11.6.1, 11.7.1, 12.3. .1.1 and 12.9.1.1. Each of the antibodies produced by the aforementioned cell lines comprises fully human IgG2 or IgG4 heavy chains with human kappa light chains. Generally, the antibodies of the invention have very high affinity, usually having a K_d from about 10 '⁹ up to about 10 '¹¹ M, when measured in solid phase or in solution.

It will be appreciated that antibodies of the invention can be expressed in cell lines other than hybridoma cell lines.

The DNA coding sequences or genomic clones for individual antibodies can be used to transform appropriate mammalian or non-mammalian host cells. Transformation can be performed by any of the known methods for introducing polynucleotides into host cells, including, but not limited to, packaging the polynucleotides into viruses (or into viral vectors) and transducing the host cells with a virus (or vector) or a transfection procedure known in the art, examples of which are illustrated in the patents USA 4,399,216, 4,912,040, 4,740,461, and 4,959,455. The transformation procedures employed will depend on the host to be transformed. Methods for introducing heterologous polynucleotides into mammalian cells are well known in the art and include, but are not limited to, dextran-mediated transfection, calcium phosphate precipitation, polybrene mediated transfection, protoplast fusion, electroporation, particle bombardment, encapsulation of polynucleotides in liposomes, peptide and peptide conjugates, and peptide conjugates. direct injection of DNA into the testicles.

Mammalian cell lines available as an expression host are well known in the art and include many immortalized cell lines available from the American Type Culture Collection (ATCC), including, but not limited to, Chinese hamster ovary (CHO), NSO cells.₀, HeLa cells, baby hamster kidney (BHK) cells, monkey kidney (COS) cells, human liver cancer cells (e.g., Hep G2), and many other cell lines. Non-mammalian cells including, but not limited to, bacterial, yeast, insect, and plant cells can be used to express recombinant antibodies. Targeted metagenesis of the antibody CH2 domain to eliminate glycosylation may be advantageous to prevent alterations in immunogenicity, pharmacokinetics, and / or effector functions as a result of non-human glycosylation. The expression methods are chosen by determining which system produces the highest expression levels and produces antibodies with constitutive CTLA-4 binding properties.

Furthermore, the expression of antibodies of the invention (or other moieties derived therefrom) in producing cell lines can be enhanced by the use of a number of known techniques. For example, the glutamine synthetase and DHFR gene expression systems are widely used approaches to enhance expression under certain conditions. High expression cell clones can be identified using conventional techniques such as limiting dilution cloning and Microdrop technology. The GS arrangement is discussed in whole or in part in connection with European Patent Nos. 0 216 846, 0 256 055 and 0 323 997 and European Patent Application No. 89303964.4.

Antibodies of the invention can also be produced transgenically by obtaining a mammal or plant that is transgenic for the immunoglobulin heavy and light chain sequences of interest and producing the antibody therefrom in a recoverable form. In conjunction with the transgenic production in mammals, antibodies can be produced and recovered from the milk of goats, cows, and other mammals. See U.S. Patent Nos. 5,827,690, 5,756,687, 5,750,172, and 5,741,957.

The antibodies of the present invention can be analyzed structurally and functionally. In conjunction with antibody structures, the kappa heavy and light chain amino acid sequences were predicted from cDNA sequences obtained from the hybridomas by RT-PCR. See Examples 3 and 4 and Figures 1-8. N-terminal antibody sequencing was also performed

PL 214 003 Β1 to confirm the results discussed in Examples 3 and 4. See Example 5 and Fig. 9. Kinetic analysis of the antibodies was performed to determine the affinity. See Example 2. The antibodies of the invention based on antibody 11.2.1 and the other antibodies described herein (particularly antibodies 4.1.1, 4.8.1 and 6.1.1) have high affinity (4.1.1: 1.63 χ 10¹⁰ l / M; 4.8.1: 3.54 χ 1O¹⁰ l / M; a 6.1.1: 7.2 χ 10⁹ l / M). In addition, the antibodies were analyzed by isoelectric focusing (IEF), reducing gel electrophoresis (SDS-PAGE), size exclusion chromatography, liquid chromatography / mass spectroscopy, and mass spectroscopy, and hybridoma production of antibodies was tested. See Example 6 and Fig. 10.

In connection with functional analysis of the antibodies of the invention, such antibodies have been shown to be potent inhibitors of CTLA-4 and its binding to ligands of the B7 family of molecules. For example, the antibodies of the invention have been shown to block CTLA-4 binding to either B7-1 or B7-2. See Example 7. Indeed, the antibodies of the invention and other antibodies disclosed herein have nanomolar and subnanomolar IC₅₀ with respect to inhibition of CTLA-4 binding to B7-1 and B7-2. Moreover, these antibodies have excellent selectivity for CTLA-4 over CD28, CD44, B7-2 or hIgG1. See Example 8. Selectivity is a ratio that reflects the degree of preferential binding of a molecule to the first factor as compared to the binding of the molecule to the second and possibly other molecules. Herein, selectivity refers to the degree of preferential binding of the antibody as compared to the binding of the antibody to other molecules such as CD28, CD44, B7-2, or hIgG1. The selectivity values ​​of the antibodies of the invention are greater than 500: 1. The antibodies of the invention have also been shown to induce or increase the expression of certain cytokines (such as IL-2 and IFN-γ) by co-culturing T cells in a T-blast cell model. See Examples 9 and 10 and Figures 12-17. Moreover, it is expected that the antibodies of the invention will inhibit tumor growth in suitable in vivo models. The design of such models is discussed in Examples 11 and 12.

The results presented herein indicate that the antibodies of the invention and the rest of the antibodies disclosed herein possess certain characteristics that make these antibodies more effective than the currently known therapeutic antibodies for CTLA-4.

In particular, the 4.1.1,4.8.1 and 6.1.1 antibodies have highly desirable properties. Their structural properties, functions, and activities provide criteria that facilitate the design or selection of additional antibodies or other molecules discussed herein. Such criteria include, but are not limited to, one or more of the following:

The ability to compete for binding with one or more antibodies of the invention;

Similar binding specificity to CTLA-4 as one or more antibodies of the invention;

Binding affinity for CTLA-4 of approximately 10 '⁹ M or greater and preferably about 10 '¹° M or more;

No cross reactivity with CTLA-4 from lower mammal species, including preferably mouse, rat and rabbit, and more preferably mouse and rat CTLA-4;

Cross-reactivity with CTLA-4 from primates, preferably CTLA-4 from rhesus macaques (Macaca mulatta) and cynomolgus (Macaca fuscata);

A selectivity for CTLA-4 over CD28, B7-2, CD44, and hIgG1 of greater than about 100: 1, and preferably about 300, 400, or 500: 1 or greater;

IC₅₀ of about 100 nM or less, and preferably 5, 4, 3, 2, 1.0.5, or 0.38 nM or less when blocking CTLA-4 binding to B7-2;

IC₅₀ of about 100 nM or less, and preferably 5, 4, 3, 2, 1.0.5, or 0.50 nM or less when blocking CTLA-4 binding to B7-1;

Increasing the production of cytokines in one or more in vitro tests:

Increasing the production of IL-2 in a T / Raji blast cell assay by about 500 pg / ml or more, preferably 750, 1000, 1500, 2000, 3000, or 3846 pg / ml or more;

Increasing the production of IFN-γ by a T / Raji blast cell assay by about 500 pg / ml or more or more, preferably 750, 1000 or 1233 pg / ml or more: or

Increasing IL-2 production in hPBMC or whole blood superantigen assay by about 500 µg / ml or greater, and preferably 750, 1000, 1200, or 1511 µg / ml or greater; In other words, it is desirable that IL-2 production be increased in the assay by about 30, 35, 40, 45, 50 percent or more relative to the control.

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Antibodies (or molecules designed or synthesized therefrom) having one or more of these properties would be expected to have similar efficacy to the antibodies described herein.

The desirable functional properties discussed above are often the result of the molecule (i.e., an antibody, antibody fragment, peptide, or small molecule) binding to and inhibiting CTLA-4 in a similar fashion to the antibody of the invention (i.e., binding to the same or a similar epitope of a CTLA molecule). -4). The molecule can be administered directly (ie, by direct administration of such molecules to the patient). Or, alternatively, the molecule may be "administered" indirectly (i.e. a peptide, or a similar agent thereof, that produces an immune response in a patient (similar to a vaccine), wherein the immune response includes the production of antibodies that bind to the same or a similar epitope, or an antibody or fragment thereof that is generated in situ upon administration of genetic material which codes for such antibodies or fragments thereof that bind to the same or a similar epitope). Thus, it will be appreciated that an epitope on CTLA-4 to which antibodies of the invention bind may be useful in the context of drug manufacturing and / or design. Negative information is also useful in drug design (ie useful is the fact that an antibody that binds to CTLA-4 does not appear to bind to an epitope that acts as a CTLA-4 inhibitor). Thus, the epitope to which the antibodies of the invention bind without leading to the desired functionality can be very useful. Also provided herein are molecules (and in particular antibodies) that bind to the same or similar epitopes as the antibodies of the invention.

In addition, we have performed some preliminary epitope mapping studies for certain antibodies, particularly antibodies 4.1.1 and 11.2.1.

As a first step, we conducted a BIAcore competition study to create a preliminary binding map between certain antibodies based on their ability to compete for CTLA-4 binding. For this purpose, CTLA-4 was bound to the BIAcore chip, the first antibody was attached to it under saturation conditions, and the subsequent binding of the second antibody to CTLA-4 was measured.

This technique makes it possible to generate a preliminary map from which antibodies of the appropriate family can be categorized.

Through this process, we have determined that certain antibodies, including the antibodies of the invention, can be divided into the following epitope categories:

<td>Category</td><td>Antibodies</td><td>Competition for CTLA-4 binding</td>

<td rowspan="2">AND</td><td>BO1M *</td><td rowspan="2">Fully free competition with each other; cross-competes with category B; some competition with category D.</td>

<td>BO2M "</td>

<td rowspan="2">B</td><td> 4.1.1</td><td rowspan="2">Fully free competition with each other; cross-competes with category A, C and D;</td>

<td> 4.13.1</td>

<td rowspan="6">C.</td><td> 6.1.1</td><td rowspan="6">Fully free competition with each other; cross-competes with category B and category D;</td>

<td> 3.1.1</td>

<td> 4.8.1</td>

<td> 11.2.1</td>

<td> 11.6.1</td>

<td> 11.7.1</td>

<td>D</td><td> 4.14.3</td><td>It cross-competes with category B and C; some competition with category A.</td>

<td rowspan="2">E.</td><td> 4.9.1</td><td rowspan="2">BIN3 blocks 4.9.1 from binding to CTLA4, but not vice versa</td>

<td>BNI3 * "</td>

(*) (**) available from Biostride (***) available from Pharmingen

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In the next step, we attempted to determine if the antibodies of the invention and other antibodies disclosed herein recognize a linear epitope on CTLA-4 under reducing and non-reducing conditions by Western blot analysis. We observed that neither of the 4.1.1, 11.7.1, 11.6.1, or 11.2.1 antibodies appeared to recognize the reduced form of CTLA-4 in Western blot analysis. Thus, it seems likely that the epitope to which each of these antibodies binds is not a linear epitope, and more likely was a conformational epitope whose structure may have been damaged under reducing conditions.

Thus, we wanted to see if we could determine which residues within the CTLA-4 molecule are essential for binding the antibodies of the invention. One of the methods we used was to perform kinetic measurements of the dissociation rates between human CTLA-4 and two highly conserved CTLA-4 primate molecules (CTLA-4 cynomolgus and marmosets). BIAcore studies have shown that antibody 4.1.1 binds human CTLA-4, cynomolgus and marmosets at the same rate. Regarding the dissociation rate (affinity), however, antibody 4.1.1 had the highest affinity (lowest dissociation rate) for human CTLA-4, a faster dissociation rate for cynomolgus CTLA-4, and the highest dissociation rate for marmoset CTLA-4. On the other hand, antibody 11.2.1 of the invention binds to human, cynomolgus and marmoset CTLA-4 at the same rate and has almost the same dissociation rate for each. These data further indicate that antibodies 4.1.1 and 11.2.1 bind to different epitopes on CTLA-4.

In order to further investigate the epitopes to which category B and C antibodies bind, we performed some studies with site-directed mutagenesis. Marmoset CTLA-4 has two important changes at residues 105 and 106 from human CTLA-4. Such differences are the change from leucine to methionine at position 105 and the change from glycine to serine at position 106. Thus, we mutated the DNA encoding human CTLA-4 to encode the mutant CTLA-4 with the L105M and G106S substitutions. The homologous exchange in CTLA-4 had no effect on the binding of the B7.2-IgG1 fusion protein. Moreover, the binding of antibody 11.2.1 was not altered. However, the molecules had a severely inhibited ability to bind to antibody 4.1.1 (similar to marmoset CTLA-4). We then mutated the DNA encoding marmoset CTLA-4 to create a CTLA-4 marmoset mutant containing the S106SG substitution. This alteration restores stable binding between the 4.1.1 antibody and the marmoset CTLA-4 mutant. In addition, we mutated the marmoset CTLA-4 cDNA to create a marmoset CTLA-4 mutant with the M105L substitution. This change partially restored the binding between the 4.1.1 antibody and the marmoset CTLA-4 mutant.

Each of the antibodies belonging to categories B to D appear to have similar functional properties and appear to have the ability to act as a potent anti-CTLA-4 therapeutic agent. Moreover, each of the molecules provides for cross competition in binding to CTLA-4. However, as will be seen from the discussion above, each of the molecules of different categories appears to bind to distinct conformational epitopes on CTLA-4.

From the foregoing, it will be concluded that the epitope data discussed above indicate that antibodies (or other molecules as discussed above) that compete with the antibodies of the invention are likely to have some therapeutic potential. Furthermore, it is expected that antibodies (or other molecules as discussed above) that cross-compete with antibodies of category B, C and / or D are likely to have some therapeutic potential. Additionally, it can be expected that antibodies (or other molecules as discussed above) that cross-compete with category B, C and / or D antibodies and which (i) do not show reduced CTLA-4 binding to marmosets (similar to the 11.2 antibody) .1) or (ii) show reduced binding to CTLA-4 marmosets (similar to antibody 4.1.1) are likely to have some therapeutic potential. Antibodies (or other molecules as discussed above) that cross-compete with categories A and E may also have some therapeutic potential. Examples

The following examples, including the experiments carried out and the results obtained, are presented for purposes of illustration only and are not intended to limit the present invention. Example 1

Preparation of hybridomas producing anti-CTLA-4 antibody

Antibodies of the invention were prepared, selected, and tested in accordance with the present Example.

Antibody production: Three distinct immunogens were produced for immunization of XenoMouse ™ mice: (i) CTLA-4-Ig fusion protein, (ii) CTLA-4 peptide, and (iii) murine lymphoma cells 300.19

PL 214 003 Β1 transfected with mutant CTLA-4 (Y201V) which is constitutively expressed on the cell surface.

(i) CTLA-4-IgG1 fusion protein: Expression vector construction:

The cDNA encoding the mature CTLA-4 extracellular domain was PCR amplified from a human thymus cDNA library (Clontech) using primers designed from the published sequence (Eur. J Immunol 18: 1901-1905 (1988)). The fragment was cloned in the correct orientation in pSR5, a Sindbis virus expression plasmid (Invitrogen), between the human oncostatin M signal peptide and the human IgG gamma 1 (IgG1) CH1 / CH2 / CH3 domains. The fusion protein does not contain a hinge domain but does contain cysteine ​​120 in the extracellular domain of CTLA-4 to form a covalent dimer. The resulting vector was named CTLA-4-IgG1 / pSR5. The sequence of the complete CTLA-4-IgG1 cDNA in the vector was confirmed for both strands. The amino acid sequence of the CTLA4-Ig protein is shown below. The mature extracellular domain for CD44 was amplified by PCR from a human lymphocyte library (Clontech) and cloned into PsinRep5 to generate a control protein with an identical IgG1 tail.

OM-CTLA4-IgG1 fusion protein:

MGVLLTORTLLSLVLALLFPSMASMAMHVAOPAVVLASSRGIASFVC

EYASPGKATEVRVTVLRQADSQVTEVCAATYMMGNELTFLDDSICT GTSSGNQVNLTIQGLRAMDTGLYICKVELMYPPPYYLGIGNGTQIY VIDPEPCPDSDLEGAPSVFLFPPKPKDTLM1SRTPEVTCWVDVSHEDPE VKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKE YKCKVSNKALPTPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCL VKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSR WQQGNVFSCSVMHEALHNHYTQKSLSLSPGK

Underline: signal peptide

Bold: CTLA-4 extracellular domain cDNA encoding the mature extracellular domain of CD28 was amplified by PCR from the Human Lymphocyte Library (Clontech) and then cloned into pCDM8 (J. Immunol. 151: 5261-71 (1993)) to generate a human fusion protein IgG1 containing both the thrombin cleavage site and the hinge regions. Marmoset, Cynomolgus and Rhesus CTLA-4 were cloned from mRNA isolated from PHA stimulated PBMCs using standard degenerate PCR techniques. Sequencing showed that the amino acid sequence for the rhesus and cynomolgus macaque was identical except for three differences from the extracellular domain of human CTLA-4 mature (S13N, I17T and L105M). The marmoset sequence showed ten amino acid differences from the extracellular domain of human CTLA4 mature (V21A, V331, A41T, A51G, 54I, S71 F, Q75K, T88M, L105M and G106S). Site-directed mutagenesis was used to obtain single mutations of all amino acids different in CTLA-4 marmosets in order to map amino acids important for antibody interactions with human CTLA-4-IgG. Mutations to map the human CTLA-IgG epitope and marmosets were generated by site-directed mutagenesis with the matchmaker site-directed mutagenesis kit (Promega). IgG fusion proteins were prepared by transient transfection of Cos7 cells and purified using standard protein A techniques. CTLA4-IgG mutant proteins were assessed for binding to antibodies by filter immunoassays and BIAcore analyzes.

Recombinant protein expression / purification

Recombinant sindbis virus was obtained by electroporation (Gibco) of hamster kidney BHK cells with in vitro transcribed CTLA-4-IgG 1 / pSRS mRNA in the SP6 system and DH-26S mRNA as described by Invitrogen. Forty-eight hours later recombinant virus was harvested and titrated for optimal expression in ovarian cells

PL 214 003 Β1 Chinese hamster (CHO-K1). CHO-K1 cells were grown in suspension in DMEM / F12 (Gibco) containing 10% heat-inactivated fetal bovine serum (Gibco), non-essential amino acids (Gibco), 4mM glutamine (Gibco), penicillin / streptomycin (Gibco), 10mM Hepes pH 7.5 (Gibco). To generate CTLA-4-IgG, CHO-K1 cells were resuspended at a concentration of 1x10⁷ cells / ml in DMEM / F12 and incubated with sindbis virus for one hour at room temperature. Cells were then diluted to 1x10⁶/ ml in DMEM / F12 containing 1% bovine IgG-free fetal bovine serum using Protein A - sepharose (Pharmacia), non-essential amino acids (Gibco), 4 mM glutamine (Gibco), 10 mM Hepes pH 7.5 (Gibco ), penicillin / streptomycin. Forty-eight hours after infection, the cells were centrifuged and the conditioned medium was harvested and protease inhibitor tablets (Boehringer Mannheim) were added to it, adjusted to pH 7.5, and filtered through a 0.2 µ filter (Nalgene). FPLC (Pharmacia) was used for affinity purification of the fusion protein using a 5 ml HiTrap protein A column (Pharmacia) at a flow rate of 10 ml / min. The column was washed with PBS 30 bed volumes and eluted with 0.1 M glycine / HCl pH 2.8 at a flow of 1 ml / min. Fractions (1 ml) were immediately neutralized to pH 7.5 with Tris pH 9. Fractions containing CTLA-4-IgG1 were identified by SDS-PAGE and then concentrated using Centriplus 50 (Amicon) before loading onto a sepharose 200 column (Pharmacia) at 1 ml / min using PBS as solvent. Fractions containing CTLA-4-IgG1 were pooled, sterile filtered through a 0.2 µ filter (Millipore), aliquoted and frozen at -80 ° C. CD44-IgG1 was expressed and purified using the same methods. CD28-IgG was purified from conditioned media from the transiently transfected Cos7 cells.

Characteristics of CTLA-4-IgG1:

Purified CTLA-4-IgG1 migrated as a single band on SDS-PAGE using Coommassie colloidal staining (Novex). Under non-reducing conditions CTLA-4-IgG1 is a dimer (100 kDa) that is reduced to a 50 kDa monomer when treated with 50 mM DTT. Determination of the amino acid sequence of the purified CTLA-4-IgG1 in solution confirmed the correct N-terminus of CTLA-4 (MHVAQPAWLAS) and that the oncostatin signal peptide was cleaved from the mature fusion protein.

CTLA-4-IgG1 bound to immobilized B7.1-IgG in a concentration dependent manner and binding was blocked by a hamster-anti-human anti-CTLA-4 antibody (BNI3: PharMingen). Sterile CTLA-4-IgG was endotoxin free and quantified by OD280 using 1.4 as the extinction coefficient. The purification yield of CTLA-4-IgG was between 0.5-3 mg / liter of CHO-K1 cells.

(ii) CTLA-4 peptide

The following CTLA-4 peptide was prepared as described below:

NH₂: MHVAQPAVVLASSRGIASFVCEYASPGKATEVRVTVLRQADSQVT

EVCAATYMMGNELTFLDDSICTGTSSGNQVNLTIQGLRAMDTGLYICK VELMYPPPYYLGIGNGTQIYV1DPEPC-CONH₂

Markings / Materials:

NMP, N-methylpyrrolidinone; TFE, 2,2,2-trifluoroethanol; DCM, dichloromethane; FMOC, Fluorenylmethoxycarbonyl. All reagents were provided by Perkin Elmer with the following exceptions: TFE, Aldrich Chemical, FMOC-PAL-PEG resin, Perseptive Biosystems. Fmoc-Arg (pMC) OH, FMOC-Asn (Trt) -OH, FMOC-Asp (tBu) -OH, FMOC-Cys (Trt) -OH, FMOCGIu (tBu) -OH, FMOC-Gln (Trt) OH, FMOC-His (Boc) -OH, FMOC-Lys (BOC) OH, FMOC-Ser (tBu) -OH, FMOC-Tr (tBu) -OH and FMOC-Tyr (tBu) -OH have been used for amino acids requiring side protecting groups .

Peptide synthesis:

Peptide synthesis was performed on a Perkin-Elmer 431 A with a UV absorbance 301nm feedback kit (Perkin-Elmer Model 759A detector). Peptide sequences were assembled onto FMOC-PAL-PEG resin using conditional double junction cycles. Forced double connections were performed on cycles 10, 11, 18, 19, 20, and 28 to 33. The resin was washed with a 50% mixture of DCM and TFE at the end of each acylation cycle, followed by blocking of unreacted amino groups with acetic anhydride in NMP. The resins were removed from the reactor at the end of cycle 49 and the reaction continued for the remainder to completion. Cleavage of the peptide from the resin was performed using

PL 214 003 Β1 of reagent K (King et al. International Journal of Protein and Peptide Research 36: 255-266 (1990)) for 6 hours on 415 mg of resin yielding 186 mg of crude CTLA-4 peptide.

Characteristics of the peptide:

Samples of 25 mg crude CTLA-4 peptide were dissolved in 5 ml of 6M guanidine HCl / 100 mM K₂AFTER₃ at pH 6.4 and eluted on a Pharmacia Hi Load Superdex 75 16/60 column (16 mm x 600 mm, bed volume 120 ml) 2 M Guanidine HCl / 100 mM K₂AFTER₃ at pH 6.4 and a rate of 2 ml / min, collecting 5 ml fractions for 180 minutes. Fractions were analyzed by loading 1.7 µl of fractions on a Laemeli NuPAGE gel, MES buffer gel and visualized using the Daichii silver staining protocol. Fractions showing a molecular weight of 12 kDa, as estimated by comparison with the molecular weight standard, were pooled and stored at 4 ° C. The pooled fractions were analyzed by UV and gel electrophoresis. Amino acid sequencing was performed by absorbing a 100 microlitre sample on a ProSorb cartridge (absorbed on a PVDF membrane). This was washed to remove buffer salts. Sequencing was performed on an Applied Biosystems 420 instrument. The expected N-terminal sequence (MHVAQPA VVLA) was observed. Immunoblotting showed that the peptide was recognized by anti-human CTLA-4 BNI3 (PharMingen). For desalination, a sample containing 648 pg of material was placed in a 3500 Da MWCO dialysis tube and dialyzed under agitation against 0.1% TFA / H₂At 4 ° C for 9 days. The entire content of the dialysis bag was lyophilized to a powder.

(iii) 300.19 cells transfected with CTLA-4 (Y201V) The full length CTLA-4 cDNA was PCR purified from a human thymus cDNA library (Stratagene) and subcloned into pIRESneo (Clontech). A CTLA-4 mutation resulting in constitutive expression on the cell surface was introduced using the MatchMaker Mutagenesis System (Promega). The tyrosine mutation, Y201, to valine inhibits the binding of the AP50 adaptin protein, which is responsible for the rapid intenalization of CTLA-4 (Chuang et al. J. Immunol. 159: 144-151 (1997)). Mycoplasma-free mouse lymphoma cells 300.19 were cultured in RPMI-1640 containing 10% fetal calf serum, non-essential amino acids, penicillin / streptomycin, 2 mM glutamine, 12.5 mM Hepes pH 7.5, and 25 µM beta-mercaptoethanol. Cells were electroporated (3x10⁶/ 0.4 ml serum free RPMI) in a 1 ml 20 µg CTLA-4-Y201V / plRESneo chamber using 200 V / 1180 pF (Gibco CeliPorator). Cells were left for 10 minutes and then 8 ml of pre-warmed RPMI medium was added. After 48 hours, cells were diluted to 0.5 x 10⁶/ ml in complete RPMI medium containing 1 mg / ml G418 (Gibco). Resistant cells were expanded and shown to express CTLA-4 on the cell surface using phycoerythrin-conjugated BNI3 antibody (PharMingen). Highly expressed cells were isolated by sterile sorting.

Immunization and hybridoma production: XenoMouse mice (8 to 10 weeks of age) were immunized subcutaneously at the base of the tail 1x10⁷ 300.19 cells that were transfected to express CTLA-4 as described above were suspended in phosphate buffered saline with complete Freund's adjuvant either (ii) subcutaneously at the base of the tail (a) 10 pg of the CTLA-4 fusion protein or (b) 10 pg of CTLA-4 peptide emulsified with complete Freund's adjuvant. In each case, the dose was repeated three or four times in incomplete Freund's adjuvant. Four days before the fusion, mice received a final injection of immunogen or cells in PBS. Spleen and / or lymph node lymphocytes from immunized mice were fused with the non-secreting mouse P3 myeloma cell line and subjected to HAT selection as previously described (Galfre. G. and Milstein, C., "Preparation of monoclonal antibodies: strategies and Procedures." Methods Enzymol. 73: 3-46 (1981)). An extensive panel of hybridomas was obtained, all of which secreted human IgG antibodies₂K or lgG₄K, as detected below) CTLA-4 specific that were recovered.

ELISA test: The ELISA test was performed to detect antigen-specific antibodies in mouse serum and hybridoma supernatants as described (Coligan et al., Chapter 2.1, "Enzyme-linked immunosorbent assays" in Current protocols in immunology (1994)) using the CTLA fusion protein -4-1g for antibody capture. For animals that are immunized with the CTLA-4-Ig fusion protein, we additionally performed an assay for non-specific reactivity against the human Ig portion of the fusion protein. This was done using ELISA plates coated with human IgG1 serving as a negative control for specificity.

The following techniques were used in a preferred ELISA assay:

ELISA plates were antigen coated at 100 µg / well in coating buffer for plates (0.1 M carbonate buffer, pH 9.6 and NaHCO₃ (MW 84) 8.4 g / l). The plates were then incubated at 4 ° C

PL 214 003 Β1 overnight. After incubation, the coating buffer is removed and the plates are blocked with a blocking buffer (0.5% BSA, 0.1% Tween 20, 0.01% thiomersal in 1 x PBS) at 200 μ w / well and incubated at room temperature for 1 hour. Alternatively, the plates are stored in a refrigerator with blocking buffer and plate sealant. Blocking buffer is removed and 50 µL / well of hybridoma supernatant, serum or other hybridoma supernatant (positive control) or HAT medium or blocking buffer (negative control) is added. Plates are incubated for 2 hours at room temperature. After incubation, the plates are washed with washing buffer (1x PBS). The detection antibody (i.e. mouse anti-human IgG2-HRP (SB, # 9070-05) for IgG2 antibodies or mouse anti-human IgG4-HRP (SB # 9200-05) for IgG4 antibodies) is added at 100 μΙ / well (mouse anti-human IgG2 -HRP at 1: 2000 dilution or mouse anti-human IgG4HRP at 1: 1000 dilution (each diluted in blocking buffer). Plates are incubated for 1 hour at room temperature. After incubation, the plates are washed with washing buffer. Then 100 pg / well of freshly prepared development solution (10 ml of stock buffer, 5 mg of OPD (o-phenylenediamine, Sigma Cat. No. P-7288) and 10 pg of 30% H) is added to the wells.₂ABOUT₂ (Sigma)). Plates are allowed to develop for 10-20 minutes until the negative control wells begin to turn color. Then, 100 pg / well stop solution (2MH₂SO₄) and the plates are read in an ELISA plate reader at 490 nm.

Measurement of the affinity constants of fully human monoclonal antibodies using BIAcore:

Affinity measurement of purified human monoclonal antibodies, Fab fragments, or hybridoma supernatants by plasmon resonance was performed on a BIAcore 2000 instrument using the general procedures described by the manufacturer.

The kinetic analysis of the antibodies was performed using the antigens immobilized on the sensor surface at low density. The fusion protein was attached to three surfaces of the sensor microplate at a density ranging from about 390 to 900 using CTLA-4-1g fusion protein at a concentration of 20 or 50 µg / ml in 10 mM sodium acetate pH 5.0 using an amine binding kit provided by manufacturer (BIAcore, Inc.). To the fourth surface of the sensor microplate, IgG1 (900 RU) was immobilized and used as a negative control for non-specific binding. Kinetic analysis was performed at a flow rate of 25 or 50 microliters per minute and dissociation rates (kd or k_ABOUTff) and associations (ka or k_he) were determined using the software provided by the manufacturer (BIA score 3.0) which enables a global calculation of the fit. Example 2

Measurement of the affinity of anti-CTLA-4 antibodies

The affinity measurement for certain antibodies is provided in the following table:

Table I.

<td></td><td colspan="5">Solid phase (BiAcore)</td>

<td>Hybridoma</td><td>Setting speed K._and(M '¹S '¹x10⁶)</td><td>Dissociation rate K._d(S '¹x10 '⁴)</td><td>The association constant Ka (1 / M) = Ka / k_dx10¹⁰</td><td>The dissociation constant K._d(M) = K._d/ kaXl O '¹⁰</td><td>Surface density [RU]</td>

<td> 1</td><td> 2</td><td> 3</td><td> 4</td><td> 5</td><td> 6</td>

<td>Moab01</td><td> 0.68</td><td> 1.01</td><td> 0.67</td><td> 1.48</td><td> 878.7</td>

<td></td><td> 0.70</td><td> 4.66</td><td> 0.15</td><td> 6.68</td><td> 504.5</td>

<td></td><td> 0.77</td><td> 6.49</td><td> 0.19</td><td> 8.41</td><td> 457.2</td>

<td></td><td> 0.60</td><td> 3.08</td><td> 0.20</td><td> 5.11</td><td> 397.8</td>

<td> 4.1.1</td><td> 1.85</td><td> 0.72</td><td> 2.58</td><td> 0.39</td><td> 878.7</td>

<td></td><td> 1.88</td><td> 1.21</td><td> 1.55</td><td> 0.64</td><td> 504.5</td>

<td></td><td> 1.73</td><td> 1.54</td><td> 1.13</td><td> 0.88</td><td> 457.2</td>

<td></td><td> 1.86</td><td> 1.47</td><td> 1.26</td><td> 0.79</td><td> 397.8</td>

PL 214 003 Β1 cont. table I.

<td> 1</td><td> 2</td><td> 3</td><td> 4</td><td> 5</td><td> 6</td>

<td> 4.8.1</td><td> 0.32</td><td> 0.07</td><td> 4.46</td><td> 0.22</td><td> 878.7</td>

<td></td><td> 0.31</td><td> 0.23</td><td> 1.33</td><td> 0.75</td><td> 504.5</td>

<td></td><td> 0.28</td><td> 0.06</td><td> 4.82</td><td> 0.21</td><td> 397.8</td>

<td> 4.14.3</td><td> 2.81</td><td> 3.04</td><td> 0.92</td><td> 1.08</td><td> 878.7</td>

<td></td><td> 2.88</td><td> 3.97</td><td> 0.73</td><td> 1.38</td><td> 504.5</td>

<td></td><td> 2.84</td><td> 6.66</td><td> 0.43</td><td> 2.35</td><td> 457.2</td>

<td></td><td> 3.17</td><td> 5.03</td><td> 0.63</td><td> 1.58</td><td> 397.8</td>

<td> 6.1.1</td><td> 0.43</td><td> 0.35</td><td> 1.21</td><td> 0.83</td><td> 878.7</td>

<td></td><td> 0.46</td><td> 0.90</td><td> 0.51</td><td> 1.98</td><td> 504.5</td>

<td></td><td> 0.31</td><td> 0.51</td><td> 0.61</td><td> 1.63</td><td> 457.2</td>

<td></td><td> 0.45</td><td> 0.79</td><td> 0.57</td><td> 1.76</td><td> 397.8</td>

<td> 3.1.1</td><td> 1.04</td><td> 0.96</td><td> 1.07</td><td> 0.93</td><td> 878.7</td>

<td></td><td> 0.95</td><td> 1.72</td><td> 0.55</td><td> 1.82</td><td> 504.5</td>

<td></td><td> 0.73</td><td> 1.65</td><td> 0.44</td><td> 2.27</td><td> 457.2</td>

<td></td><td> 0.91</td><td> 2.07</td><td> 0.44</td><td> 2.28</td><td> 397.8</td>

<td> 4.9.1</td><td> 1.55</td><td> 13.80</td><td> 0.11</td><td> 8.94</td><td> 878.7</td>

<td></td><td> 1.43</td><td> 19.00</td><td> 0.08</td><td> 13.20</td><td> 504.5</td>

<td></td><td> 1.35</td><td> 20.50</td><td> 0.07</td><td> 15.20</td><td> 397.8</td>

<td> 4.10.2</td><td> 1.00</td><td> 2.53</td><td> 0.39</td><td> 2.54</td><td> 878.7</td>

<td></td><td> 0.94</td><td> 4.30</td><td> 0.22</td><td> 4.55</td><td> 504.5</td>

<td></td><td> 0.70</td><td> 5.05</td><td> 0.14</td><td> 7.21</td><td> 457.2</td>

<td></td><td> 1.00</td><td> 5.24</td><td> 0.19</td><td> 5.25</td><td> 397.8</td>

<td> 2.1.3</td><td> 1.24</td><td> 9.59</td><td> 0.13</td><td> 7.72</td><td> 878.7</td>

<td></td><td> 1.17</td><td> 13.10</td><td> 0.09</td><td> 11.20</td><td> 504.5</td>

<td></td><td> 1.11</td><td> 13.00</td><td> 0.09</td><td> 11.70</td><td> 397.8</td>

<td> 4.13.1</td><td> 1.22</td><td> 5.83</td><td> 0.21</td><td> 4.78</td><td> 878.7</td>

<td></td><td> 1.29</td><td> 6.65</td><td> 0.19</td><td> 5.17</td><td> 504.5</td>

<td></td><td> 1.23</td><td> 7.25</td><td> 0.17</td><td> 5.88</td><td> 397.8</td>

As will be seen, the antibodies show high affinity and binding constants. Example 3

Structure of anti-CTLA-4 antibodies

In the following discussion, structural information regarding the antibodies of the invention is provided.

In order to analyze the antibody structures, we cloned genes encoding heavy and light chain fragments from some hybridomas. Cloning and sequencing were performed as follows:

PL 214 003 Β1 mRNA poly (A)⁺ was isolated using the Fast-Track kit (Invitrogen) at approximately 2 × 10⁵ hybridoma cells derived from an immunized XenoMouse mouse. The production of cDNA using random primers was monitored by PCR. Primers specific for the variable regions of the human V family_H. or Vk (Marks et al., "Oligonucleotide primers for polymerase chain reaction amplification of human immunoglobulin variable genes and design of family-specific oligonucleotide probes." Eur. J. Immunol. 21: 985-991 (1991)) or a universal primer for human V_H., MG-30 (CAGGTGCAGCTGGAGCAGTCIGG) was used in conjunction with primers specific for the human Cyl constant region (MG-40d; 5'-GCTGAGGGAGTAGTCCTGAGGA-3 ') or the Ck constant region (ΙίκΡ2; as previously described in Green et al., 1994). The sequences of the heavy chain and kappa derived transcripts from the hybridomas were obtained by direct sequencing of PCR products made from poly (A⁺) using the primers described above. The PCR products were also cloned into pCRII using the TA cloning kit (Invitrogen) and both strands were sequenced using the Prism colored terminator sequencing kit and the ABI 377 sequencing machine. All sequences were analyzed by aligning the "V BASE sequence directory" (Tomlinson et al. ., MRC Center for Protein Engineering, Cambridge, UK) using MacVector and Geneworks software.

In addition, each of the antibodies 4.1.1,4.8.1, 11.2.1 and 6.1.1 were sequenced along their entire length. For such sequencing, poly (A) mRNA was isolated⁺ with about 4χ10⁶ hybridoma cells using the mRNA Direct Kit (Dynal). mRNA was reverse transcribed using oligo-dT (18) and the Advantage for RT / PCR kit (Clonetech). A database for the variable region (V Base) was used to design amplification primers beginning at the ATG start site of the heavy chain of the DP50 gene (5'-TATCTAAGCTTCTAGACTCGACCGCCACCATGGAGTTTGGGCTGATG-3 ') and to the stop codon of the IgG2 constant region_i5t.TTCTCTGATCAGAATTCCTATCATTTACCCGGAGACAGGGAGAGCT-3 ').

On the 5 'side of the ATG start site, the optimal Kozak sequence (ACCGCCACC) was added.

The same method was used to design a primer for the ATG start site of the kappa chain of the A27 gene (5'-TCTTCAAGCTTGCCCGGGCCCGCCACCATGGAAACCCCAGCGCA-3 ') and for the kappa constant region stopcodon (5'-TTCTTTGATCAGAATTCTCACTAACTAACTAACCTCCAGATGTTTCCAGATCAGAATTCTCACTAACTAACTCCTCC.

cDNA 012 was cloned with the ATG start site primer (5'-TCTTCAAGCTTGCCCGGGCCCGCCACCATGGACATGAGGGTCCCCG T-3) and the kappa constant region stop codon as described above. The heavy chain DNA was also cloned as genomic constructs by site-directed mutagenesis to add a NheI site to the end of the J variable domain and subcloning of the NheI fragment containing the genomic IgG2 CH1 / Hinge / CH2 / CH3 regions. Point mutations to generate a NheI site do not alter the amino acid sequence from germline. The primer pairs were used to amplify the DNA using the Advantage High Fidelity PCR kit (Clonetech). The sequence of the PCR products was obtained by direct sequencing using a colored terminator sequencing kit and an ABI sequencing device. The PCR product was cloned into pEE glutamate synthetase (Lonza) mammalian expression vectors and the three clones were sequenced to confirm somatic mutations. For each clone, the sequence was checked from two strands in at least three reactions. Non-glycosylated antibody 4.1.1 was generated by site-directed mutagenesis of N294Q in the CH2 domain. Recombinant antibodies were generated by transient transfection of Cos7 cells in IgG-free FCS and purified using standard protein A sepharose techniques. Stable transfectants were produced by electroporation of mouse NSO cells and selection on glutamine free medium. Recombinant antibodies 4.1.1

PL 214 003 Β1 with or without glycosylation showed identical specificity and affinity for CTLA-4 in the in vitro ELISA and BIAcore assays.

Gene utilization analysis

The following table shows the gene utilization demonstrated by the selected hybridoma antibody clones:

Table II

Use of light and heavy chain

<td rowspan="2">Clone</td><td colspan="3">Heavy chain</td><td colspan="2">Kappa light chain</td>

<td>VH</td><td>D</td><td>JH</td><td>VK</td><td>JK</td>

<td> 4.1.1</td><td>DP-50</td><td>DIR4 or DIR3</td><td>JH4</td><td>A27</td><td>JK1</td>

<td> 4.8.1</td><td>DP-50</td><td> 7-27</td><td>JH4</td><td>Λ27</td><td>JK4</td>

<td> 4.14.3</td><td>DP-50</td><td> 7-27</td><td>JH4</td><td>A27</td><td>JK3</td>

<td> 6.1.1</td><td>DP-50</td><td>DIR5 or DIR5rc</td><td>JH4</td><td>A27</td><td>JK3</td>

<td> 3.1.1</td><td>DP-50</td><td> 3-3</td><td>JH6</td><td> 012</td><td>JK3</td>

<td> 4.10.2</td><td>DP-50</td><td> 7-27</td><td>JH4</td><td>A27</td><td>JK3</td>

<td> 2.1.3</td><td>DP-65</td><td> 1-26</td><td>JH6</td><td>Α10 / A26</td><td>JK4</td>

<td> 4.13.1</td><td>DP-50</td><td> 7-27</td><td>JH4</td><td>A27</td><td>JK3</td>

<td> 11.2.1</td><td>DP-50</td><td>D1-26</td><td>JH6</td><td> 012</td><td>JK3</td>

<td> 11.6.1</td><td>DP-50</td><td>D2-2 or D4</td><td>JH6</td><td> 012</td><td>JK3</td>

<td> 11.7.1</td><td>DP-50</td><td>D3-22 or D21-9</td><td>JH4</td><td> 012</td><td>JK3</td>

<td> 12.3.1.1</td><td>DP-50</td><td>D3-3 or DXP4</td><td>A17</td><td>A17</td><td>JK1</td>

<td> 12.9.1.1</td><td>DP-50</td><td>D6-19</td><td>A3 / A19</td><td>A3 / A19</td><td>JK4</td>

<td> 4.9.1</td><td>DP-47</td><td>5-24 and or 6-19</td><td>L5</td><td>L5</td><td>JK1</td>

As will be seen, antibodies have been produced with a strong tendency to use DP-50 heavy chain variable regions. The DP-50 gene is also referred to as the V gene family_H. 3-33. Only one antibody that was selected on the basis of CTLA-4 binding and preliminary in vitro functional analyzes showed a heavy chain utilization other than DP-50. This clone, 2.1.3, uses the heavy chain region and the IgG4 isotype. The DP-65 gene is also referred to as the V gene family_H. 4-31. On the other hand, clone 4.9.1, which has the DP-47 heavy chain variable region, binds to CTLA-4 but does not inhibit binding to B7-1 or B7-2. In XenoMouse mice, there are over 30 distinct functional variable heavy chain genes with which it produces antibodies. The tendency therefore indicates a preferred binding of the antigen-antigen interaction motif with respect to the combined antigen binding properties and functional activity.

Analysis by mutation

As will be appreciated, gene utilization analysis only provides a general understanding of the structure of an antibody. Since B cells of XenoMouse animals randomly produce VDJ heavy chain or VJ kappa light chain transcripts, many secondary processes occur, including but not limited to somatic hypermutations, N-terminal additions, and CDR3 extension. See, for example, Mendez et al. Nature Genetics 15: 146-156 (1997) and US Patent Application Serial No. 08 / 759,620 filed December 3, 1996. Thus, to further investigate the structure of the antibody, the predicted amino acid sequences of the antibodies were obtained from the cDNA obtained from these clones. In addition, N-terminal amino acid sequences were determined by protein sequencing.

PL 214 003 Β1

Figure 1 shows the nucleotide sequences and predicted amino acid sequences of the kappa heavy and light chain clones 4.1.1 (Figure 1A), 4.8.1 (Figure 1B), 4.14.3 (Figure 1C), 6.1.1 (Figure 1). 1D), 3.1.1 (Fig. 1E), 4.10.2 (Fig. 1F), 2.1.3 (Fig. 1G), 4.13.1 (Fig. 1H), 11.2.1 (Fig. 11), 11.6 .1 (Fig. 1J), 11.7.1 (Fig. 1K), 12.3.1.1 (Fig. 1L), and 12.9.1.1 (Fig. 1M). In Figures 1A, 1B, and 1D, the extended sequences for antibodies 4.1.1,4.8.1 and 6.1.1 were obtained by cloning the full length cDNA as described above. In these figures, the sequences of the signal peptide (or their encoding bases) are shown in bold and the sequences used in the 5'PCR reaction are underlined.

Figure 2 shows the alignment of the predicted heavy chain amino acid sequences from clones 4.1.1, 4.8.1, 4.14.3, 6.1.1, 3.1.1, 4.10.2, 4.13.1, 11.2.1, 11.6.1, 11.7. 1, 12.3.1.1 and 12.9.1.1 and the germline amino acid sequence of DP-50 (3-33). Differences between DP-50 germline sequence and clone sequence are shown in bold. The figure also shows the positions of the CDR1, CDR2 and CDR3 sequences as a shade.

Figure 3 shows the alignment of the predicted amino acid sequences of the heavy chain of clone 2.1.3 and the germline amino acid sequence of DP-65 (4-31). Differences between the DP-65 germline sequence and the clone sequence are shown in bold. The figure also shows the positions of the CDR1, CDR2 and CDR3 sequences of the antibody as underlined.

Figure 4 shows the alignment of the predicted kappa light chain amino acid sequences of clones 4.1.1,4.8.1,4.14.3, 6.1.1,4.10.2 and 4.13.1 and the germline A27 amino acid sequence. Differences between the A27 germline sequence and the clone sequence are shown in bold. The figure also shows the positions of the CDR1, CDR2 and CDR3 sequences of the antibody as underlined. The observed deletions in the CDR1 of clones 4.8.1,4.14.3 and 6.1.1 are marked as "0".

Figure 5 shows the alignment of the predicted kappa light chain amino acid sequences of clones 3.1.1, 11.2.1, 11.6.1, and 11.7.1 and the germline 012 amino acid sequences. Differences between the germline 012 sequence and the clone sequence are shown in bold. The figure also shows the positions of the CDR1, CDR2 and CDR3 sequences of the antibody as underlined.

Figure 6 shows the alignment of the predicted amino acid sequences of the kappa light chain of clone 2.1.3 and the germline A10 / A26 amino acid sequence. Differences between the A10 / A26 germline sequence and the clone sequence are shown in bold. The figure also shows the positions of the CDR1, CDR2 and CDR3 sequences of the antibody as underlined.

Figure 7 shows the alignment of the predicted amino acid sequences of the kappa light chain of clone 12.3.1 and the germline A17 amino acid sequence. Differences between the A17 germline sequence and the clone sequence are shown in bold. The figure also shows the positions of the CDR1, CDR2 and CDR3 sequences of the antibody as underlined.

Figure 8 shows the alignment of the predicted amino acid sequences of the kappa light chain of clone 12.9.1 and the germline A3 / A19 amino acid sequence. Differences between the A3 / A19 germline sequence and the clone sequence are shown in bold. The figure also shows the positions of the CDR1, CDR2 and CDR3 sequences of the antibody as underlined.

Figure 22 shows a series of additional nucleotide and amino acid sequences of the following anti-CTLA-4 antibody chains:

4.1.1:

full-length heavy chain 4.1.1 (cDNA sequence 22 (a), genomic 22 (b) and amino acid 22 (c));

full-length heavy chain 4.1.1, non-glycosylated (cDNA sequence 22 (d) and amino acid 22 (e));

light chain 4.1.1 (cDNA sequence 22 (f) and amino acid 22 (g));

4.8.1:

full length heavy chain 4.8.1 (cDNA sequence 22 (h) and amino acid 22 (i));

4.8.1 light chain (cDNA sequence 22 (j) and amino acid sequence 22 (k));

6.1.1:

full length heavy chain 6.1.1 (cDNA sequence 22 (1) and amino acid sequence 22 (m)); 6.1.1 light chain (cDNA sequence 22 (n) and amino acid 22 (o));

11.2.1:

full length 11.2.1 heavy chain (cDNA sequence 22 (p) and amino acid 22 (q)); and the light chain 11.2.1 (cDNA22 (r) and amino acid 22 (s) sequence).

PL 214 003 Β1

Signal peptide sequences are shown in bold and enlarged letters. The open reading frames in the full-length genome sequence (Fig. 22 (b)) are underlined. The mutations introduced to produce a non-glycosylated heavy chain of 4.1.1 and the resulting change (N294O) is shown as double underline and bold (cDNA (Fig. 22 (b)) and amino acid (Fig. 22 (c)) sequence).

Example 4

Analysis of amino acid substitutions in heavy and light chains

From Fig. 2, which shows the alignment of the predicted heavy chain amino acid sequences from clones 4.1.1,4.8.1,4.14.3, 6.1.1,3.1.1,4.10.2, 4.13.1, 11.2.1, 11.6.1 , 11.7.1, 12.3.1.1 and 12.9.1.1 and the germline amino acid sequence of DP-50 (3-33), an interesting relationship emerges. In addition to the tendency to use the DP-50 heavy chain in most clones, there is relatively limited hypermutation in the antibodies relative to the DP-50 germline. For example, clones 3.1.1 and 11.2.1 have no mutation. Furthermore, mutations in other clones are usually conservative changes, involving substitutions with amino acids with similar properties to those of germline amino acids. Mutations within many CDR1 and CDR2 sequences are especially conserved in nature. Three of the heavy chains shown in Figures 2, 4.10.2, 4.13.1, 4.14.3 are clearly derived from one recombination event (i.e. come from an identical germinal center) and have an almost identical sequence. If treated as a single sequence, then among the 10 different antibodies containing the DP-50 heavy chain, there are 3 positions in CDR1 and CDR2 where a non-polar residue is replaced with another non-polar residue, 12 where a polar uncharged residue is replaced with a different polar residue. an uncharged residue and 1 in which a polar residue is replaced with a different polar residue. In addition, there are two positions where two residues that are very similar structurally, glycine and alanine, are replaced with each other. The only incompletely conserved mutations include 3 substitutions of polar charged polar residues with uncharged residues, and 1 substitution of a non-polar residue with a polar residue.

The light chains of these antibodies are derived from 5 different Vk genes. The A27 gene is the most highly represented and is the source of 6 different light chains. The comparison of these 6 sequences shows two properties worth mentioning. First, three of them, 4.8.1,4.14.3 and 6.1.1, contain deletions of one or two residues in CDR1, which is a rare event. Second, there is a strong bias against the germline serine at position six in CDR3 as manifested in that this serine has been replaced in each sequence. This suggests that the serine at this position is incompatible with CTLA-4 binding.

It will be appreciated that many of the substitutions identified above are in close proximity to or within the CDRs. Such substitutions may have some effect on the binding of the antibody to the CTLA-4 molecule. Moreover, such substitutions will have a significant effect on the affinity of the antibodies.

Example 5

Analysis of the N-terminal amino acid sequence of the antibodies

In order to further verify the composition and structure of the antibodies of the invention identified above, we sequenced some of the antibodies using a Perkin-Elmer sequencer. Both the heavy and light chains of the antibodies were isolated and purified using preparative gel electrophoresis and filter electrotransfer techniques, and then directly sequenced as described in Example 6. Most of the heavy chain sequences were blocked at the amino terminus. Thus, such antibodies were treated first with pyroglutamate aminopeptidase and then sequenced.

The result of this experiment is shown in Fig. 9. Fig. 9 also shows the molecular weights of the heavy and light chains as determined by mass spectroscopy (MALDI). Example 6

Additional characteristics of antibodies

Figure 10 provides some additional information regarding the characteristics of some antibodies. In Figure 10A, data on clones 3.1.1,4.1.1,4.8.1,4.10.2, 4.14.3 and 6.1.1 are collected. Data shown are: concentration, isoelectric focusing (IEF), SDS-PAGE, size exclusion chromatography, mass spectroscopy (MALDI) and N-terminal light chain sequences.

PL 214 003 Β1

Basically the data was obtained as described below:

Materials and methods

Protein concentration was determined at 280 nm by UV scan (200-350 nm), with an absorbance of 1.58 units at 280 nm corresponding to 1 mg / ml.

SDS-PAGE was performed on a Novex NuPAGE electrophoresis system using a 10% NuPAGE gel and MES electrophoresis buffer. The samples were prepared by diluting 3: 1 in 4 x NuPAGE (+/-) beta-mercaptoethanol sample buffer, heated and ~ 5 Lg of protein was applied to the gel. The gel was then stained with Brilliant Blue R staining solution (Sigma cat. # B6529) and the molecular weight was estimated by comparing the stained bands with the "Perfect Protein Markers" (Novagen cat # 69149-3).

For N-terminal sequencing, samples were run on a NuPAGE gel as above, transferred to a Pro Biot filter (Applied Biosystems), and then stained with Coomassie Blue R-250 staining solution. The stained protein bands were excised and subjected to sequence analysis by automated Edman degradation on an Applied Biosystems 494 Procise HT Sequencer.

Isoelectric focusing (JEF) was performed using Pharmacia IEF 3-9 pHast gels (Cat # 17-0543-01). The samples were diluted in 10% glycerol to -0.8 mg / ml and 1 μΙ was applied to the gel and then stained with silver. Estimation of pi was done by comparing the stained bands to the broad range (pH 3-10) IEF standards (Pharmacia cat # 17-0471-01).

Size exclusion chromatography (SEC) was performed in phosphate buffered saline (PBS) on a Pharmacia SMART system using a Superdex 75 PC 3.2 / 30 column. Molecular weight estimation was done by comparing the hold time peak with the gel hold time.

For FACS studies, human peripheral T cells were prepared and stimulated for 48 hours. T cells were washed once, resuspended in 1x10 FACS buffer⁶ cells / 100 µ and stained for CD3 surface expression with 10 µΙ anti-CD3-FITC antibodies (Immunotech, Marseille, France) for 30 minutes at room temperature. Cells were washed twice, then fixed, permeabilized (membrane permeability increased) (Fix and Penn, Caltag) and stained for surface CTLA-4 expression with 10 µl anti-CD1S2-PE (Pharmingen). Flow cytometry was performed using a Becton Dickinson FACSort. Quadrants were determined by isotype control antibody analysis (Caltag).

As discussed above, anti-CTLA-4 antibodies have been shown to possess some potent immune system modulating properties. The following experiments were performed to determine whether the antibodies of the invention possess such activities. In general, the experiments were designed to test the ability of antibodies to inhibit the interaction between CTLA-4 and B7 molecules, to be selective for CTLA-4 against B7 and CD28 molecules, and to promote the production of cytokines for T cells, including but not limited to IL-2 expression. and / or IFN-γ. In addition, studies were undertaken on the cross-reactivity of antibodies with certain human tissues and CTLA-4 molecules from other species (e.g., mice and primates).

Example 7

Competition ELISA: Inhibition of CTLA-4 / B7-1 or B7-2 Interactions by Antibodies

An in vitro assay was performed to determine whether the antibodies of the invention had the ability to inhibit CTLA-4 binding to either B7-1 or B7-2. As will be appreciated, antibodies that have the ability to inhibit CTLA-4 binding to B7 molecules will be candidates for regulating the immune response through the CTLA-4 pathway. The following materials and methods were used in the test:

Materials and Methods nM B7.1-Ig (G1) or B7.2-Ig (G1) (Repligen, Inc. Needham, MA) in Dulbecco PBS were used to coat 96-well MaxiSorp plates (Nunc, Denmark, # 439454) and incubated overnight at 4 ° C. On day 2, B7-Ig was removed and plates were blocked with 1% BSA plus 0.05% Tween-20 in D-PBS for two hours. Plates were washed 3X with wash buffer (0.05% Tween-20 in D-PBS). Antibodies at appropriate concentrations and CTLA-4-Ig (G4) (0.3 nM final concentration) (Repligen, Inc. Needham, MA) was pre-mixed for 15 minutes then added to B7-1g coated plates (60 µΙ total volume) and incubated at room temperature for 1.5 hours. Plates were washed 3X and 50 μΙ of a 1 in 1000 dilution of HRP-labeled mouse anti-human was added

PL 214 003 Β1 of IgG4 antibodies (Zymed, San Francisco, CA, # 05-3820) and incubated for 1 hour at room temperature (RT). Plates were washed 3X and 50 μΙ of TMB Microwell peroxidase substrate (Kirkegaard &amp; Peuy, Gaithersburg, MD, # 50-76-04) was added and incubated at RT for 20 minutes, then 50 μΙ of 1N H was added to the plate.₂SO₄. Plates were read at 450 nm using a Molecular Device plate reader (Sunnyvale, CA). All plates were tested in duplicate. Peak signal was defined as CTLA-4-Ig binding in the absence of antibody. Nonspecific binding was defined in the absence of CTLA-4-1g and test antibody.

The results of this test are shown in Tables 111A and IIIB. In Table IHA, the results are shown for the various antibodies. In Table IIIB, the results comparing antibody 4.1.1 to antibody 11.2.1 of the invention from a separate experiment are shown.

Table IIIA

<td>Clone CTLA-4-Ig</td><td>Isotype</td><td>CTLA4 / B7.2 Comp. ELISA IC $ o (nM)</td><td>CTLA4 / B7.1 Comp. ELISA IC $ o (nM)</td>

<td>CT3.1.1</td><td>lgG2</td><td>0.45 ± 0.07 (n = 3)</td><td>0.63 ± 0.10 (n = 2)</td>

<td>CT4.1.1</td><td>lgG2</td><td>0.38 ± 0.06 (n = 5)</td><td>0.50 ± 0.05 (n = 2)</td>

<td>CT4.8.1</td><td>igG2</td><td>0.57 ± 0.03 (n = 3)</td><td>0.17 ± 0.28 (n = 2)</td>

<td>CT4.9.1</td><td>igG2</td><td>Non-competitive (n = 3)</td><td>Non-competitive (n = 2)</td>

<td>CT4.10.2</td><td>igG2</td><td>1.50 ± 0.37 (n = 3)</td><td>3.39 + 0.31 (n = 2)</td>

<td>CT4.13.1</td><td>igG2</td><td>0.49 ± 0.05 (n = 3)</td><td>0.98 ± 0.11 (n = 2)</td>

<td>CT4.14.3</td><td>igG2</td><td>0.69 ± 0.11 (n = 3)</td><td>1.04 ± 0.15 (n = 2)</td>

<td>CT6.1.1</td><td>igG2</td><td>0.39 ± 0.06 (n = 3)</td><td>0.67 ± 0.07 (n = 2)</td>

Table IIIB

<td>Clone CTLA-4-Ig</td><td>Isotype</td><td>CTLA4 / B7.2 ELISA Comp IC50 (nM)</td><td>CTLA4 / B7.1 ELISA Comp IC50 (nM)</td>

<td>CT4.1.1</td><td>lgG2</td><td>0.55 ± 0.08 (n = 4)</td><td>0.87 ± 0.14 (n = 2)</td>

<td>CT11.2.1</td><td>lgG2</td><td>0.56 ± 0.05 (n = 4)</td><td>0.81 ± 0.24 (n = 2)</td>

Example 8

Ratio of antibody selectivity for CTLA-4 versus CD28 or B7-2

Another in vitro test was performed to determine the selectivity of the CTLA-4 antibodies against either CD28 or B7-2. In connection with the experiments, the following materials and methods were used:

Selectivity ELISA for CTLA-4: Materials and Methods

A 96-well FluroNUNC plate (Nunc Cat # 475515) was precoated with the four antigens: CTLA-4 / Ig, CD44 / Ig, CD28 / Ig and B7.2 / Ig (in-house prepared antigens). The plates were coated with antigens overnight at + 4 ° C at a concentration of 1 µg / ml and 100 µΙ / well in 0.1 M sodium phosphate, pH 9.6. The plate was then washed three times with PBST (PBS + 0.1% Tween-20) using a NUNC plate washer. The plates were blocked with PBST + 0.5% BSA at a concentration of 150 µM / well. Plates were incubated at RT for 1 hour and then washed three times with PBST. The anti-CTLA-4 antibodies were then diluted to a concentration of 1 µg / ml and added to the plate. Plates were incubated at RT for 1 hour and then washed three times with PBST. The plates containing the antibodies were then treated with 100 µ / well anti-human IgG2-HRP (Southern Biotech cat # 9070-05) at a dilution of 1: 4000. In addition, one row was treated with anti-human IgG (Jackson Cat No. 209-035-088) to normalize the precoating. Antibody was diluted 1: 5000 and added at 100 µΙ / well. One row was also treated with anti-human CTLA-4-HRP (Pharmingen Cat No. 345815 / HRP conjugated) as a positive control. This antibody will be used

PL 214 003 Β1 was counted as block dilution to 0.05 pg / ml. Plates were incubated at RT for 1 hour and then washed three times with PBST. Chemiluminescent LBA substrate (Pierce) was added at a concentration of 100 µl / well and the plates were incubated on a plate shaker for 5 min. The plates were then read using a fluorescence detector for 2 min exposure.

IGEN CTLA-4-Ig selectivity binding assay: Materials and Methods

Dynabeads M-450 beads (Dynal AS, Oslo, Norway # 140.02) were washed 3X with Na phosphate buffer, pH 7.4 and resuspended in phosphate buffer. 1.0 pg CTLA-4-Ig (G1), 1.0 pg CD28-Ig (G1) or 1.0 to 3.0 pg B7.2-Ig (G1) (Repligen, Inc. Needham, MA) were added to 100 µl of beads and incubated overnight on a rotary shaker at 4 ° C. On the second day, the beads were washed 3X with 1% BSA plus 0.05% Tween-20 in Dulbecco's PBS and blocked for 30 minutes. The beads were diluted 1 to 10 with blocking buffer and 25 µl of the coated beads were added to 12x75 mm polypropylene tubes. All samples were tested in duplicate. 50 µl of test antibody (1 µg / ml final concentration) or blocking buffer was added to the tubes and incubated for 30 minutes on an Origen 1.5 Analyzer carousel (IGEN International, Inc., Gaithersburg, MD) at RT with 100 rpm agitation. 25 µl of ruthenylated mouse anti-human IgG1, IgG2, or IgG4 (Zymed, Inc. San Francisco, CA # 05-3300, 05-3500 and 05-3800) was added to the tubes (final concentration of 3 µg / ml in a total volume of 100 µl) . The tubes were incubated for 30 minutes at RT on the carousel with agitation at 100 rpm. 200 µl of Origen Buffer (IGEN International, Inc., Gaithersburg, MD # 402-050-03) was added to each tube briefly and the tubes were counted on the Origen Analyzer and ECL units (electrochemiluminescence) were determined for each tube. The normalization factor was determined to correct for differences in binding of the fusion proteins to Dynabeads and the ECL units were corrected for non-specific binding before calculating selectivity ratios.

The results of these tests are shown in Tables IV A and IVB.

Table IVA

<td>Clone</td><td>Izo type</td><td>CTLA4.CD28 ELISA</td><td>CTLA4 / B7.2 ELISA</td><td>CTLA4 / CD44 ELISA</td><td>CTLA4 / CD28 IGEN</td><td>CTLA4 / B7.2 IGEN</td>

<td> 3.1.1</td><td>lgG2</td><td>&gt; 500: 1 (n = 3)</td><td>&gt; 500: 1 (n = 3)</td><td>&gt; 500: 1 (n = 3)</td><td>&gt; 500: 1 (n = 2)</td><td>&gt; 500: 1 (n = 1) 195: 1 (n = 1)</td>

<td> 4.1.1</td><td>lgG2</td><td>&gt; 500: 1 (n = 3)</td><td>&gt; 5001 (n = 2) 485: 1 (n = 1)</td><td>&gt; 500: 1 (n = 3)</td><td>&gt; 500: 1 (n = 1) 261: 1 (n = 1)</td><td>&gt; 500: 1 (n = 1) 107: 1 (n = 1)</td>

<td> 4.8.1</td><td>igG2</td><td>&gt; 500: 1 (n = 3)</td><td>&gt; 500: 1 (n = 2) 190: 1 (n = 1)</td><td>&gt; 500: 1 (n = 3</td><td>&gt; 500: 1 (n = 2)</td><td>&gt; 500: 1 (n = 2)</td>

<td> 4.9.1</td><td>igG2</td><td>&gt; 500: 1 (n = 2) 244: 1 (n = 1)</td><td>&gt; 500: 1 (n = 2) 33: 1 (n = 1)</td><td>&gt; 500: 1 (n = 3)</td><td>&gt; 500: 1 (n = 1)</td><td>&gt; 500: 1 (n = 1)</td>

<td> 4.10.2</td><td>igG2</td><td>&gt; 500: 1 (n = 3)</td><td>&gt; 500: 1 (n = 3)</td><td>&gt; 500: 1 (n = 3)</td><td>&gt; 500: 1 (n = 1)</td><td>&gt; 500: 1 (n = 1)</td>

<td> 4.13.1</td><td>igG2</td><td>&gt; 500: 1 (n = 2) 46: 1 (n = 1)</td><td>&gt; 500: 1 (n = 3)</td><td>&gt; 500: 1 (n = 3)</td><td>&gt; 500: 1 (n = 1) 329: 1 (n = 1)</td><td>&gt; 500: 1 (n = 2)</td>

<td> 4.14.3</td><td>igG2</td><td>&gt; 500: 1 (n = 2) 80: 1 (n = 1)</td><td>&gt; 500: 1 (n = 2) 10: 1 (n = 1)</td><td>&gt; 500: 1 (n = 2) 126: 1 (n = 1)</td><td>&gt; 413: 1 (n = 1)</td><td>&gt; 234: 1 (n = 1)</td>

<td> 6.1,1</td><td>igG2</td><td>&gt; 500: 1 (n = 2) 52: 1 (n = 1)</td><td>&gt; 500: 1 (n = 3)</td><td>&gt; 500: 1 (n = 3)</td><td>&gt; 500: 1 (n = 2)</td><td>&gt; 500: 1 (n = 2)</td>

Table IVB

<td>Clone</td><td>Isotope</td><td>CTLA4 / CD28 ELISA</td><td>CTLA4 / B7.2 ELISA</td><td>CTLA4 / hlgG ELISA</td>

<td> 4.1.1</td><td>lgG2</td><td>-&gt; 500: 1 (n = 3;</td><td>&gt; 500: 1 (n = 2)</td><td>&gt; 500: 1 (n = 3)</td>

<td> 11.2.1</td><td>lgG2</td><td>&gt; 500: 1 (n = 3)</td><td>&gt; 500: 1 (n = 3)</td><td>&gt; 500: 1 (n = 3)</td>

Example 9

The human model of T cell signaling

In order to further define the activity of antibodies in acting as regulators of immune activity, we have developed some T-cell assays to quantify the potentiation

PL 214 003 Β1 of IL-2 production by T cells when CTLA-4 signal is blocked by antibodies. The following materials and methods were used in connection with the experiments:

Materials and methods

Freshly isolated T cells were obtained by using Histopaque (Sigma, St. Louis, MO # A-70543) and iT-kwik (Lympho-Kwik, One Lambda, Canoga Park, CA, # LK-50-T) and stimulated with PHA (1 µg / ml) (purified phytohaemagglutinin, Murex Diagnostics Ltd. Dartford, England, #HA 16) in medium (RPMI 1640 containing L-glutamine, essential amino acids MEM, penicillin, streptomycin, 25 mM Hepes and 10% FBS) at a concentration of 1 χ 10⁶ cells / ml and incubated at 37 ° C for two days. The cells were then washed and diluted in medium to 2x10⁶ cells / ml. Raji cells (Burkitt's lymphoma, Human ATCC #: CCL 86 Class 1.1 American Type Culture Collection Rockville, MD) were then treated with mitomycin C (Sigma St. Louis, MO, # M-4287) (25 μ 25 / ml) for 1 hour at 37 ° C. Raji cells were then washed 4X in PBS and resuspended at a concentration of 2x10⁶ cells / ml. Human T blasts (5x10⁵/ ml), Raji cells (5x10⁵/ ml) and anti-CTLA-4 antibodies or an isotype matching control antibody at various concentrations were added to 96-well microtiter plates and the plates were incubated at 37 ° C for 72 hours. The total volume per well was 200 µL. Seventy-two hours after stimulation, the plates were centrifuged and the supernatant removed and frozen for further determination of IL-2 (Quantikine IL-2 ELISA kit, R&amp;D Systems, Minneapolis, MN, # D2050) and IFN-γ (Quantikine IFN-g ELISA kit) , R&amp;D Systems). Cytokine enhancement was defined as the difference between the cytokine levels in cultures containing the anti-CTLA-4 blocking mAb versus the isotype matched control antibody. For flow cytometry experiments, Raji cells were washed 1x with FACS buffer (PBS buffer containing 2% heat-inactivated FOS, 0.025% sodium azide). Cell pellets were resuspended in FACS buffer at a concentration of 1x10⁶ cells / 100 µL and incubated with 10 µL anti-CD80-PE (Becton Dickinson, San Jose, CA) or anti-CD86-PE (Pharmingen, San Diego, CA) for 30 minutes at room temperature. The cells were then washed twice and resuspended in 1 ml of FACS buffer. Flow cytometry was performed using a Becton Dickinson FACSort. Histogram markers were established by analyzing appropriate isotype control antibodies (Caltag, Burlingame, CA).

Overall, we have developed an assay that can be used to rapidly determine the increase in IL-2 secretion by T cells. As will be appreciated, stimulation of T cells is dependent on B7 and CD28. Moreover, washed T-blasts do not produce IL-2 and Raji cells do not produce detectable amounts of IL-2 even after LPS or PWM stimulation. However, in combination, T blast cells co-cultured with Raji cells can model B7, CTLA-4, and CD28 signaling and thus the effect of antibodies on such signaling can be assessed.

Figure 11 shows the expression of B7-1 and B7-2 on Raji cells using anti-CD80PE and anti-CD86-PE mAbs using flow cytometry (FASC) as described in Example 6.

Fig. 12 shows the concentration-dependent enhancement of IL-2 production in the T cell blast / Raji cell assay induced by blocking anti-CTLA-4 antibodies (BNI3 (PharMingen) and antibodies 4.1.1,4.8.1 and 6.1.1).

Fig. 13 shows the concentration-dependent enhancement of IFN-γ production in a blast T / Raji cell assay induced by blocking anti-CTLA-4 antibodies (BNI3, 4.1.1,4.8.1 and 6.1.1) (same donor T cells) .

Fig. 14 shows the mean enhancement of IL-2 production in T cells from 6 donors induced by blocking anti-CTLA-4 antibodies in the T blastocyte / Raji cell assay. Interestingly, the mAb, CT4.9.1, binds to CTLA-4 but does not block binding to B7. Thus, simple binding to CTLA-4 is alone insufficient to provide a functional antibody of the invention.

Figure 15 shows the mean enhancement of IFN-γ production in T cells from 6 donors induced by blocking anti-CTLA-4 antibodies in the T cell blast / Raji cell assay.

Fig. 19 shows the comparison between antibodies 4.1.1 and 11.2.1 of the invention at a concentration of 30 µg / ml in the 72 hour T / Raji blast cells assay as described in Example 9 and the superantigen assay described in Translation 10.

Fig. 20 shows the concentration-dependent enhancement of IL-2 production in T blastocytes / Raji cells induced by anti-CTLA-4 antibodies 4.1.1 and 11.2.1.

Table IVc below provides information on the mean enhancement and extent of cytokine response production by Raji and SEA assays. Each of the experiments included in the results is based on an antibody dose of 30 µg / ml and measured at 72 hours. The numbers of donors used in these experiments as well as the responses are shown.

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Table IVc

<td>Test</td><td>has b</td><td>Cytokine</td><td>Average gain pg / ml</td><td>SEM</td><td>Amplification range pg / ml</td><td>n</td><td>Donor's answer</td>

<td>T blastocytes / Raji</td><td> 4.1.1</td><td>IL-2</td><td> 3329</td><td> 408</td><td>0 to 8861</td><td> 42</td><td>19z21</td>

<td>T blastocytes / Raji</td><td> 4.1.1</td><td>IFNs</td><td> 3630</td><td> 980</td><td>600 to 13939</td><td> 17</td><td>13 of 13</td>

<td>T blastocytes / Raji</td><td> 11.2.1</td><td>IL-2</td><td> 3509</td><td> 488</td><td>369 to 6424</td><td> 18</td><td>14 of 14</td>

<td>SEA (PBMC)</td><td> 4.1.1</td><td>IL-2</td><td> 2800</td><td> 312</td><td>330 to 6699</td><td> 42</td><td>17 of 17</td>

<td>SEA (PMBC)</td><td> 11.2.1</td><td>IL-2</td><td> 2438</td><td> 366</td><td>147 to 8360</td><td> 25</td><td>15 of 15</td>

<td>SEA (whole blood)</td><td> 4.1.1</td><td>IL-2</td><td> 6089</td><td> 665</td><td>-168 to 18417</td><td> 46</td><td>15 of 17</td>

<td>SEA (whole blood)</td><td> 11.2.1</td><td>IL-2</td><td> 6935</td><td> 700</td><td>-111 to 11803</td><td> 25</td><td>12 of 14</td>

Example 10

Model of signaling by human T cells

We developed a second cell assay to quantify the potentiation of IL-2 production by T cells when the CTL-4 signal was blocked by antibodies.

The following materials and methods were used in connection with the experiments: Materials and methods

Human PBMCs were prepared using Accuspin. Microtiter plates were coated with anti-CD3 antibody (Ieu4, Becton Dickinson) (60 ng / ml) and incubated for 2 hours at 37 ° C. hPBMC was added to the wells at a concentration of 200,000 cells per well. Staphylcoccus Enterotoxin A (SEA) (Sigma) was added to the wells at a concentration of 100 ng / ml. Antibodies were added to the wells, usually at a concentration of 30 µg / ml. Cells were then stimulated for 48, 72 or 96 hours. Plates were centrifuged at appropriate time points and supernatants were collected from the wells. Subsequently, the supernatants were checked for IL-2 production using an ELISA assay (R&amp;D Systems).

The results of these experiments are shown in Figs. 16, 17 and 21.

In Figure 16, the induction of IL-2 production in hPBMC from 5 donors was measured 72 hours after stimulation. Fig. 17 shows the results of whole blood measurements analyzing the differences in induction of IL-2 production in blood from 3 donors measured at 72 and 96 hours after SEA stimulation.

In Fig. 21, the IL-2 production enhancement for whole blood from 2 donors was measured 72 hours after stimulation.

Example 11

Animal model of tumor

We developed an animal tumor model to analyze anti-mouse-CTLA-4 antibodies in vivo for their tumor growth inhibitory activity. In this model, fibrosarcoma was cultured in mice, and the animals were treated with anti-mouse-CTLA-4 antibodies. The materials and methods for model development are presented below:

Materials and methods

Female A / J mice (6-8 weeks old) were injected subcutaneously on the dorsal side of the neck with 0.2 ml of SaIN tumor cells (1x10⁶) (Baskar 1995). Anti-mouse CTLA-4 antibody or isotype matched control (PharMingen, San Diego, CA, 200 µg / animal) was injected intraperitoneally on days 0, 4, 7 and 14 after tumor cell injection. Tumor measurements were made over a period of 3-4 weeks of experiments using a Starrett SPC Plus electronic calibration device (Athol, MA) and tumor size was expressed as tumor covered area (mm²).

Fig. 18 shows the inhibition of tumor growth by an anti-mouse CTLA-4 antibody in a murine fibrosarcoma model. As shown in Fig. 18, animals treated with anti-CTLA-4 show a reduction in tumor growth compared to animals treated with an isotype control antibody. Thus, anti-mouse CTLA-4 mAbs are capable of inhibiting the growth of fibrosarcoma in a mouse tumor model.

PL 214 003 Β1

Antibodies that cross-react with mouse CTLA-4 would be expected to behave similarly in this model. However, antibodies of the invention that were checked for cross-reactivity did not show it with mouse CTLA-4.

Example 12

Animal model of tumor

A mouse xenograft model in SCID mice was designed to further investigate antibody activity against established tumors and their metastases. In this model, SCID mice transplanted with human T cells, implanted with patient-derived non-small cell lung (NSCL) or colorectal carcinoma (CC) cells, are provided. . Implants are made into the gonadal fat pads of SCTD mice. The tumor is allowed to grow and then removed. Mice develop a human type tumor and metastasize to the liver. Such a model is described in (Bumpers et al. J. Surgical Res. 61: 282-288 (1996).

Antibodies described herein are expected to inhibit the growth of tumors arising from such mice.

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Sequence list &lt;110&gt; ABGENIK, INC.

PFIZ-ER INC.

&lt;120&gt; Monoclonal antibody against CTLA-4 &lt;130&gt; ΑΒΧ-PFl PCT &lt;140&gt; PCT / US99 / 30895 &lt;141&gt; 1999-12-23 &lt;150&gt; 60 / 113,647 &lt;151&gt; 1999-12-23 &lt;160&gt; 147 &lt;170&gt; Patent Ver. 2.1 &lt;210&gt; 1 &lt;211&gt; 463 &lt;212&gt; PRT &lt;213&gt; Homo sapiens

<td colspan="5"> &lt;400&gt; 1</td><td rowspan="2">Cheese</td><td colspan="2" rowspan="2">Trp Val</td><td colspan="2" rowspan="2">Phe Leu 10</td><td rowspan="2">Val</td><td rowspan="2">Ala</td><td rowspan="2">Leu</td><td rowspan="2">Leu</td><td rowspan="2">Arg 15</td><td rowspan="2">Gly</td>

<td>Underworld</td><td>Glu</td><td colspan="2">Phe Gly</td><td>Leu 5</td>

<td>Val</td><td>Gin</td><td>Cys</td><td>Gin</td><td>Val</td><td>Gin</td><td>Leu</td><td>Val</td><td>Glu</td><td>Cheese</td><td>Gly</td><td>Gly</td><td>Gly</td><td>Val</td><td>Val</td><td>Gin</td>

<td></td><td></td><td></td><td> 20</td><td></td><td></td><td></td><td></td><td> 25</td><td></td><td></td><td></td><td></td><td> 30</td><td></td><td></td>

<td>Pro</td><td>Gly</td><td>Arg</td><td>Cheese</td><td>Leu</td><td>Arg</td><td>Leu</td><td>Cheese</td><td>Cys</td><td>Val</td><td>Ala</td><td>Cheese</td><td>Gly</td><td>Phe</td><td>Thr</td><td>Phe</td>

<td></td><td></td><td> 35</td><td></td><td></td><td></td><td></td><td> 40</td><td></td><td></td><td></td><td></td><td> 45</td><td></td><td></td><td></td>

<td>Cheese</td><td>Cheese</td><td>His</td><td>Gly</td><td>Underworld</td><td>His</td><td>Trp</td><td>Val</td><td>Arg</td><td>Gin</td><td>Ala</td><td>Pro</td><td>Gly</td><td>Lys</td><td>Gly</td><td>Leu</td>

<td></td><td> 50</td><td></td><td></td><td></td><td></td><td> 55</td><td></td><td></td><td></td><td></td><td> 60</td><td></td><td></td><td></td><td></td>

<td>Glu</td><td>Trp</td><td>Val</td><td>Ala</td><td>Val</td><td>How much</td><td>Trp</td><td>Tyr</td><td>Asp</td><td>Gly</td><td>Arg</td><td>Asn</td><td>Lys</td><td>Tyr</td><td>Tyr</td><td>Ala</td>

<td> 65</td><td></td><td></td><td></td><td></td><td> 70</td><td></td><td></td><td></td><td></td><td> 75</td><td></td><td></td><td></td><td></td><td> 80</td>

<td>Asp</td><td>Cheese</td><td>Val</td><td>Lys</td><td>Gly</td><td>Arg</td><td>Phe</td><td>Thr</td><td>How much</td><td>Cheese</td><td>Arg</td><td>Asp</td><td>Asn</td><td>Cheese</td><td>Lys</td><td>Asn</td>

<td></td><td></td><td></td><td></td><td> 85</td><td></td><td></td><td></td><td></td><td> 90</td><td></td><td></td><td></td><td></td><td> 95</td><td></td>

<td>Thr</td><td>Leu</td><td>Phe</td><td>Leu</td><td>Gin</td><td>Underworld</td><td>Asn</td><td>Cheese</td><td>Leu</td><td>Arg</td><td>Ala</td><td>Glu</td><td>Asp</td><td>Thr</td><td>Ala</td><td>Val</td>

<td></td><td></td><td></td><td> 100</td><td></td><td></td><td></td><td></td><td> 105</td><td></td><td></td><td></td><td></td><td> 110</td><td></td><td></td>

<td>Tyr</td><td>Tyr</td><td>Cys</td><td>Ala</td><td>Arg</td><td>Gly</td><td>Gly</td><td>His</td><td>Phe</td><td>Gly</td><td>Pro</td><td>Phe</td><td>Asp</td><td>Tyr</td><td>Trp</td><td>Gly</td>

<td></td><td></td><td> 115</td><td></td><td></td><td></td><td></td><td> 120</td><td></td><td></td><td></td><td></td><td> 125</td><td></td><td></td><td></td>

<td>Gin</td><td>Gly</td><td>Thr</td><td>Leu</td><td>Val</td><td>Thr</td><td>Val</td><td>Cheese</td><td>Cheese</td><td>Ala</td><td>Cheese</td><td>Thr</td><td>Lys</td><td>Gly</td><td>Pro</td><td>Cheese</td>

<td></td><td> 130</td><td></td><td></td><td></td><td></td><td> 135</td><td></td><td></td><td></td><td></td><td> 140</td><td></td><td></td><td></td><td></td>

<td>Val</td><td>Phe</td><td>Pro</td><td>Leu</td><td>Ala</td><td>Pro</td><td>Cys</td><td>Cheese</td><td>Arg</td><td>Cheese</td><td>Thr</td><td>Cheese</td><td>Glu</td><td>Cheese</td><td>Thr</td><td>Ala</td>

<td> 145</td><td></td><td></td><td></td><td></td><td> 150</td><td></td><td></td><td></td><td></td><td> 155</td><td></td><td></td><td></td><td></td><td> 160</td>

<td>Ala</td><td>Leu</td><td>Gly</td><td>cys</td><td>Leu</td><td>Val</td><td>Lys</td><td>Asp</td><td>Tyr</td><td>Phe</td><td>Pro</td><td>Glu</td><td>Pro</td><td>Val</td><td>Thr</td><td>Val</td>

<td></td><td></td><td></td><td></td><td> 165</td><td></td><td></td><td></td><td></td><td> 170</td><td></td><td></td><td></td><td></td><td> 175</td><td></td>

PL 214 003 Β1

<td>Cheese</td><td>Trp</td><td>Asn</td><td>Cheese 180</td><td>Gly</td><td>ΑΣ3</td><td>Leu</td><td>Thr</td><td>Cheese 185</td><td>Gly</td><td>Val</td><td>His</td><td>Thr</td><td>Phe 190</td><td>Pro</td><td>Ala</td>

<td>Val</td><td>Leu</td><td>Gin</td><td>Cheese</td><td>Cheese</td><td>Gly</td><td>Leu</td><td>Tyr</td><td>Cheese</td><td>Leu</td><td>Cheese</td><td>Cheese</td><td>Val</td><td>Val</td><td>Thr</td><td>Val</td>

<td></td><td></td><td> 195</td><td></td><td></td><td></td><td></td><td> 200</td><td></td><td></td><td></td><td></td><td> 205</td><td></td><td></td><td></td>

<td>Pro</td><td>Cheese</td><td>Cheese</td><td>Asn</td><td>Phe</td><td>dy</td><td>Thr</td><td>Gin</td><td>Thr</td><td>Tyr</td><td>Thr</td><td>Cys</td><td>Asn</td><td>Val</td><td>Asp</td><td>His</td>

<td></td><td> 210</td><td></td><td></td><td></td><td></td><td> 215</td><td></td><td></td><td></td><td></td><td> 220</td><td></td><td></td><td></td><td></td>

<td>Lys</td><td>Pro</td><td>Cheese</td><td>Asn</td><td>Thr</td><td>Lys</td><td>Val</td><td>Asp</td><td>Lys</td><td>Thr</td><td>Val</td><td>Glu</td><td>Arg</td><td>Lys</td><td>Cys</td><td>Cys</td>

<td> 225</td><td></td><td></td><td></td><td></td><td> 230</td><td></td><td></td><td></td><td></td><td> 235</td><td></td><td></td><td></td><td></td><td> 240</td>

<td>Val</td><td>Glu</td><td>Cys</td><td>Pro</td><td>Pro</td><td>Cys</td><td>Pro</td><td>Ala</td><td>Pro</td><td>Pro</td><td>Val</td><td>Ala</td><td>Gly</td><td>Pro</td><td>Cheese</td><td>Val</td>

<td></td><td></td><td></td><td></td><td> 245</td><td></td><td></td><td></td><td></td><td> 250</td><td></td><td></td><td></td><td></td><td> 255</td><td></td>

<td>Phe</td><td>Leu</td><td>Phe</td><td>Pro</td><td>Pro</td><td>Lys</td><td>Pro</td><td>Lys</td><td>Asp</td><td>Thr</td><td>Leu</td><td>Underworld</td><td>How much</td><td>Cheese</td><td>Arg</td><td>Thr</td>

<td></td><td></td><td></td><td> 260</td><td></td><td></td><td></td><td></td><td> 265</td><td></td><td></td><td></td><td></td><td> 270</td><td></td><td></td>

<td>Pro</td><td>Glu</td><td>Val</td><td>Thr</td><td>Cys</td><td>Val</td><td>Val</td><td>Val</td><td>Asp</td><td>Val</td><td>Cheese</td><td>His</td><td>Glu</td><td>Asp</td><td>Pro</td><td>Glu</td>

<td></td><td></td><td> 275</td><td></td><td></td><td></td><td></td><td> 280</td><td></td><td></td><td></td><td></td><td> 285</td><td></td><td></td><td></td>

<td>Val</td><td>Gin</td><td>Phe</td><td>Asn</td><td>Trp</td><td>Tyr</td><td>Val</td><td>Asp</td><td>dy</td><td>Val</td><td>Glu</td><td>Val</td><td>His</td><td>Asn</td><td>Ala</td><td>Lys</td>

<td></td><td> 290</td><td></td><td></td><td></td><td></td><td> 295</td><td></td><td></td><td></td><td></td><td> 300</td><td></td><td></td><td></td><td></td>

<td>Thr</td><td>Lys</td><td>Pro</td><td>Arg</td><td>Glu</td><td>Glu</td><td>Gin</td><td>Phe</td><td>Asn</td><td>Cheese</td><td>Thr</td><td>Phe</td><td>Arg</td><td>Val</td><td>Val</td><td>Cheese</td>

<td> 305</td><td></td><td></td><td></td><td></td><td> 310</td><td></td><td></td><td></td><td></td><td> 315</td><td></td><td></td><td></td><td></td><td> 320</td>

<td>Val</td><td>Leu</td><td>Thr</td><td>Val</td><td>Val</td><td>His</td><td>Gin</td><td>Asp</td><td>Trp</td><td>Leu</td><td>Asn</td><td>Gly</td><td>Lys</td><td>Glu</td><td>Tyr</td><td>Lys</td>

<td></td><td></td><td></td><td></td><td> 325</td><td></td><td></td><td></td><td></td><td> 330</td><td></td><td></td><td></td><td></td><td> 335</td><td></td>

<td>Cys</td><td>Lys</td><td>Val</td><td>Cheese</td><td>Asn</td><td>Lys</td><td>Gly</td><td>Leu</td><td>Pro</td><td>Ala</td><td>Pro</td><td>How much</td><td>Glu</td><td>Lys</td><td>Thr</td><td>How much</td>

<td></td><td></td><td></td><td> 340</td><td></td><td></td><td></td><td></td><td> 345</td><td></td><td></td><td></td><td></td><td> 350</td><td></td><td></td>

<td>Cheese</td><td>Lys</td><td>Thr</td><td>Lys</td><td>Gly</td><td>Gin</td><td>Pro</td><td>Arg</td><td>Glu</td><td>Pro</td><td>Gin</td><td>Val</td><td>Tyr</td><td>Thr</td><td>Leu</td><td>Pro</td>

<td></td><td></td><td> 355</td><td></td><td></td><td></td><td></td><td> 360</td><td></td><td></td><td></td><td></td><td> 365</td><td></td><td></td><td></td>

<td>Pro</td><td>Cheese</td><td>Arg</td><td>G1 u</td><td>Glu</td><td>Underworld</td><td>Thr</td><td>Lys</td><td>Asn</td><td>Gin</td><td>Val</td><td>Cheese</td><td>Leu</td><td>Thr</td><td>Cys</td><td>U ^ 5 U</td>

<td></td><td> 370</td><td></td><td></td><td></td><td></td><td> 375</td><td></td><td></td><td></td><td></td><td> 380</td><td></td><td></td><td></td><td></td>

<td>Val</td><td>Lys</td><td>Gly</td><td>Phe</td><td>Tyr</td><td>Pro</td><td>Cheese</td><td>Asp</td><td>How much</td><td>Ala</td><td>Vał</td><td>Glu</td><td>Trp</td><td>Glu</td><td>Cheese</td><td>Asn</td>

<td> 385</td><td></td><td></td><td></td><td></td><td> 390</td><td></td><td></td><td></td><td></td><td> 395</td><td></td><td></td><td></td><td></td><td> 400</td>

<td>Gly</td><td>Gin</td><td>Pro</td><td>Glu</td><td>Asn</td><td>Asn</td><td>Tyr</td><td>Lys</td><td>Thr</td><td>Thr</td><td>Pro</td><td>Pro</td><td>Underworld</td><td>Leu</td><td>Asp</td><td>Cheese</td>

<td></td><td></td><td></td><td></td><td> 405</td><td></td><td></td><td></td><td></td><td> 410</td><td></td><td></td><td></td><td></td><td> 415</td><td></td>

<td>Asp</td><td>Gly</td><td>Cheese</td><td>Phe</td><td>Phe</td><td>Leu</td><td>Tyr</td><td>Cheese</td><td>Lys</td><td>Leu</td><td>Thr</td><td>Val</td><td>Asp</td><td>Lys</td><td>Cheese</td><td>Arg</td>

<td></td><td></td><td></td><td> 420</td><td></td><td></td><td></td><td></td><td> 425</td><td></td><td></td><td></td><td></td><td> 430</td><td></td><td></td>

<td>Trp</td><td>Gin</td><td>Gin</td><td>Gly</td><td>Asn</td><td>Val</td><td>Phe</td><td>Cheese</td><td>Cys</td><td>Cheese</td><td>Val</td><td>Underworld</td><td>His</td><td>Glu</td><td>Ala</td><td>Leu</td>

<td></td><td></td><td> 43 5</td><td></td><td></td><td></td><td></td><td> 440</td><td></td><td></td><td></td><td></td><td> 445</td><td></td><td></td><td></td>

<td>His</td><td>Asn</td><td>His</td><td>_Tyr</td><td>Thr</td><td>Gin</td><td>Lys</td><td>Cheese</td><td>Leu</td><td>Cheese</td><td>Leu</td><td>Cheese</td><td>Pro</td><td>Gly</td><td>Lys</td><td></td>

<td></td><td> 450</td><td></td><td></td><td></td><td></td><td> 455</td><td></td><td></td><td></td><td></td><td> 4 60</td><td></td><td></td><td></td><td></td>

&lt;210&gt; 2 &lt;211&gt; 464

PL 214 003 Β1 &lt;212&gt; PRT &lt;213&gt; Homo sapiens

<td colspan="16"> &lt;400&gt; 2</td>

<td>Underworld 1</td><td>Glu</td><td>Phe</td><td>Gly</td><td>Leu 5</td><td>Cheese</td><td>Trp</td><td>Val</td><td>Phe</td><td>Leu 10</td><td>Val</td><td>Ala</td><td>Leu</td><td>Leu</td><td>Arg 15</td><td>Gly</td>

<td>Val</td><td>Gin</td><td>Cys</td><td>Gin</td><td>Val</td><td>Gin</td><td>Leu</td><td>Val</td><td>Glu</td><td>Cheese</td><td>Gly</td><td>Gly</td><td>Gly</td><td>Val</td><td>Val</td><td>Gin</td>

<td></td><td></td><td></td><td> 20</td><td></td><td></td><td></td><td></td><td> 25</td><td></td><td></td><td></td><td></td><td> 30</td><td></td><td></td>

<td>Pro</td><td>Gly</td><td>Arg</td><td>Cheese</td><td>Leu</td><td>Arg</td><td>Leu</td><td>Cheese</td><td>Cys</td><td>Thr</td><td>Ala</td><td>Cheese</td><td>Gly</td><td>Phe</td><td>Thr</td><td>Phe</td>

<td></td><td></td><td> 35</td><td></td><td></td><td></td><td></td><td> 40</td><td></td><td></td><td></td><td></td><td> 4 5</td><td></td><td></td><td></td>

<td>Cheese</td><td>Asn</td><td>Tyr</td><td>Gly</td><td>Underworld</td><td>His</td><td>Trp</td><td>Val</td><td>Arg</td><td>Gin</td><td>Ala</td><td>Pro</td><td>Gly</td><td>Lys</td><td>Gly</td><td>Leu</td>

<td></td><td> 50</td><td></td><td></td><td></td><td></td><td> 55</td><td></td><td></td><td></td><td></td><td> 60</td><td></td><td></td><td></td><td></td>

<td>Glu</td><td>Trp</td><td>Val</td><td>Ala</td><td>Val</td><td>Tl _ώ</td><td>Trp</td><td>Tyr</td><td>Asp</td><td>Gly</td><td>Cheese</td><td>Asn</td><td>^L.ys</td><td>His</td><td>Tyr</td><td>Giy</td>

<td> 65</td><td></td><td></td><td></td><td></td><td> 70</td><td></td><td></td><td></td><td></td><td> 75</td><td></td><td></td><td></td><td></td><td> 80</td>

<td>Asp</td><td>Cheese</td><td>Val</td><td>Lys</td><td>Gly</td><td>Arg</td><td>Phe</td><td>Thr</td><td>How much</td><td>Cheese</td><td>Cheese</td><td>Asp</td><td>Asn</td><td>Cheese</td><td>Lys</td><td>Asn</td>

<td></td><td></td><td></td><td></td><td> 85</td><td></td><td></td><td></td><td></td><td> 90</td><td></td><td></td><td></td><td></td><td> 95</td><td></td>

<td>Thr</td><td>Leu</td><td>Tyr</td><td>Leu</td><td>Gin</td><td>Underworld</td><td>Asn</td><td>Cheese</td><td>Leu</td><td>Arg</td><td>Al a</td><td>Glu.</td><td>Asp</td><td>Thr</td><td>Ala</td><td>Val</td>

<td></td><td></td><td></td><td> 100</td><td></td><td></td><td></td><td></td><td> 105</td><td></td><td></td><td></td><td></td><td> 110</td><td></td><td></td>

<td>Tyr</td><td>Tyr</td><td>Cys</td><td>Ala</td><td>Arg</td><td>Gly</td><td>Glu</td><td>Arg</td><td>Leu</td><td>Gly</td><td>Cheese</td><td>Tyr</td><td>Phe</td><td>Asp</td><td>Tyr</td><td>Trp</td>

<td></td><td></td><td> 115</td><td></td><td></td><td></td><td></td><td> 120</td><td></td><td></td><td></td><td></td><td> 125</td><td></td><td></td><td></td>

<td>Gly</td><td>Gin</td><td>Gly</td><td>Thr</td><td>Leu</td><td>Val</td><td>Thr</td><td>Val</td><td>Cheese</td><td>Cheese</td><td>Ala</td><td>Cheese</td><td>Thr</td><td>Lys</td><td>Gly</td><td>Pro</td>

<td></td><td> 130</td><td></td><td></td><td></td><td></td><td> 135</td><td></td><td></td><td></td><td></td><td> 140</td><td></td><td></td><td></td><td></td>

<td>Cheese</td><td>Val</td><td>Phe</td><td>Pro</td><td>Leu</td><td>Ala</td><td>Pro</td><td>Cys</td><td>Cheese</td><td>Arg</td><td>Cheese</td><td>Thr</td><td>Cheese</td><td>Glu</td><td>Cheese</td><td>Thr</td>

<td> 145</td><td></td><td></td><td></td><td></td><td> 150</td><td></td><td></td><td></td><td></td><td> 155</td><td></td><td></td><td></td><td></td><td> 160</td>

<td>Ala</td><td>Ala</td><td>Leu</td><td>Gly</td><td>Cys</td><td>Leu</td><td>Val</td><td>Lys</td><td>Asp</td><td>Tyr</td><td>Phe</td><td>Pro</td><td>Glu</td><td>Pro</td><td>Val</td><td>Thr</td>

<td></td><td></td><td></td><td></td><td> 165</td><td></td><td></td><td></td><td></td><td> 170</td><td></td><td></td><td></td><td></td><td> 17 5</td><td></td>

<td>Val</td><td>Cheese</td><td>Trp</td><td>As n</td><td>Cheese</td><td>Gly</td><td>Ala</td><td>Leu</td><td>Thr</td><td>Cheese</td><td>Gly</td><td>Val</td><td>His</td><td>Thr</td><td>Phe</td><td>Pro</td>

<td></td><td></td><td></td><td> 180</td><td></td><td></td><td></td><td></td><td> 185</td><td></td><td></td><td></td><td></td><td> 190</td><td></td><td></td>

<td>Al a</td><td>Va 1</td><td>Leu</td><td>Gin</td><td>Cheese</td><td>Cheese</td><td>Gly</td><td>Leu</td><td>Tyr</td><td>Cheese</td><td>Leu</td><td>Cheese</td><td>Cheese</td><td>Val</td><td>Val</td><td>Thr</td>

<td></td><td></td><td> 195</td><td></td><td></td><td></td><td></td><td> 200</td><td></td><td></td><td></td><td></td><td> 205</td><td></td><td></td><td></td>

<td>a 1</td><td>Pro</td><td>Cheese</td><td>Cheese</td><td>Asn</td><td>Phe</td><td>Gly</td><td>Thr</td><td>Gin</td><td>Thr</td><td>Tyr</td><td>Thr</td><td>Cys</td><td>Asn</td><td>Val</td><td>Asp</td>

<td></td><td> 210</td><td></td><td></td><td></td><td></td><td> 215</td><td></td><td></td><td></td><td></td><td> 220</td><td></td><td></td><td></td><td></td>

<td>His</td><td>Lys</td><td>Pro</td><td>Cheese</td><td>Asn</td><td>Thr</td><td>Lys</td><td>Val</td><td>Asp</td><td>Lys</td><td>Thr</td><td>Val</td><td>Glu</td><td>Arg</td><td>Lys</td><td>Cys</td>

<td> 225</td><td></td><td></td><td></td><td></td><td> 230</td><td></td><td></td><td></td><td></td><td> 235</td><td></td><td></td><td></td><td></td><td> 240</td>

<td>Cys</td><td>Val</td><td>Glu</td><td>Cys</td><td>Pro</td><td>Pro</td><td>Cys</td><td>Pro</td><td>Al a</td><td>Pro</td><td>Pro</td><td>Val</td><td>Ala</td><td>Gly</td><td>Pro</td><td>Cheese</td>

<td></td><td></td><td></td><td></td><td> 245</td><td></td><td></td><td></td><td></td><td> 250</td><td></td><td></td><td></td><td></td><td> 255</td><td></td>

<td>Val</td><td>Phe</td><td>Leu</td><td>Phe</td><td>Pro</td><td>Pro</td><td>Lys</td><td>Pro</td><td>Lys</td><td>Asp</td><td>Thr</td><td>Leu</td><td>Underworld</td><td>How much</td><td>Cheese</td><td>Arg</td>

<td></td><td></td><td></td><td> 260</td><td></td><td></td><td></td><td></td><td> 2 65</td><td></td><td></td><td></td><td></td><td> 270</td><td></td><td></td>

<td>Thr</td><td>Pro</td><td>Glu</td><td>Val</td><td>Thr</td><td>Cys</td><td>Va 1</td><td>Val</td><td>Val</td><td>Asp</td><td>Val</td><td>Cheese</td><td>His</td><td>Glu</td><td>Asp</td><td>Pro</td>

<td></td><td></td><td> 275</td><td></td><td></td><td></td><td></td><td> 280</td><td></td><td></td><td></td><td></td><td> 285</td><td></td><td></td><td></td>

PL 214 003 Β1

<td>Glu</td><td>Val 290</td><td>Gin</td><td>Phe</td><td>Asn</td><td>Trp</td><td>Tyr 2 9b</td><td>Val</td><td>Asp</td><td>Gly</td><td>Val</td><td>Glu 300</td><td>Val</td><td>His</td><td>Asn</td><td>Ala</td>

<td>Lys</td><td>Tir n</td><td>Lys</td><td>Pro</td><td>Arg</td><td>Glu</td><td>Glu</td><td>Gin</td><td>Phe</td><td>Asn</td><td>Cheese</td><td>Thr</td><td>Phe</td><td>Arg</td><td>Val</td><td>Val</td>

<td> 305</td><td></td><td></td><td></td><td></td><td> 310</td><td></td><td></td><td></td><td></td><td> 315</td><td></td><td></td><td></td><td></td><td> 320</td>

<td>Cheese</td><td>val</td><td>Leu</td><td>Thr</td><td>Val</td><td>Val</td><td>His</td><td>Gin</td><td>Asp</td><td>Trp</td><td>Leu</td><td>Asn</td><td>Gly</td><td>Lys</td><td>Glu</td><td>Tyr</td>

<td></td><td></td><td></td><td></td><td> 325</td><td></td><td></td><td></td><td></td><td> 330</td><td></td><td></td><td></td><td></td><td> 335</td><td></td>

<td>Lys</td><td>Cys</td><td>Lys</td><td>Val</td><td>Cheese</td><td>Asn</td><td>Lys</td><td>Gly</td><td>Leu</td><td>Pro</td><td>Ala</td><td>Pro</td><td>Background</td><td>Glu</td><td>Lys</td><td>Thr</td>

<td></td><td></td><td></td><td> 340</td><td></td><td></td><td></td><td></td><td> 345</td><td></td><td></td><td></td><td></td><td> 350</td><td></td><td></td>

<td>How much</td><td>Cheese</td><td>Lys</td><td>Tłir</td><td>Lys</td><td>Gly</td><td>Gin</td><td>Pro</td><td>Arg</td><td>Glu</td><td>Pro</td><td>Gin</td><td>Val</td><td>Tyr</td><td>Thr</td><td>Leu</td>

<td></td><td></td><td> 355</td><td></td><td></td><td></td><td></td><td> 360</td><td></td><td></td><td></td><td></td><td> 365</td><td></td><td></td><td></td>

<td>Pro</td><td>Pro</td><td>Cheese</td><td>Arg</td><td>Glu</td><td>Glu</td><td>Underworld</td><td>Thr</td><td>Lys</td><td>Asn</td><td>Gin</td><td>Val</td><td>Cheese</td><td>Leu</td><td>Thr</td><td>Cys</td>

<td></td><td> 370</td><td></td><td></td><td></td><td></td><td> 375</td><td></td><td></td><td></td><td></td><td> 380</td><td></td><td></td><td></td><td></td>

<td>Leu</td><td>val</td><td>Lys</td><td>Gly</td><td>Phe</td><td>f £ y £.</td><td>Pro</td><td>C! a, y</td><td>Asp</td><td>How much</td><td>Ala</td><td>Val</td><td>Glu</td><td>Trp</td><td>Glu</td><td>Cheese</td>

<td> 38 5</td><td></td><td></td><td></td><td></td><td> 390</td><td></td><td></td><td></td><td></td><td> 395</td><td></td><td></td><td></td><td></td><td> 400</td>

<td>Asn</td><td>Gly</td><td>Gin</td><td>Pro</td><td>Glu</td><td>Asn</td><td>Asn</td><td>Tyr</td><td>Lys</td><td>Thr</td><td>Thr</td><td>Pro</td><td>Pro</td><td>Underworld</td><td>Leu</td><td>Asp</td>

<td></td><td></td><td></td><td></td><td> 405</td><td></td><td></td><td></td><td></td><td> 410</td><td></td><td></td><td></td><td></td><td> 415</td><td></td>

<td>Cheese</td><td>Asp</td><td>Gly</td><td>Cheese</td><td>Phe</td><td>Phe</td><td>Leu</td><td>Tyr</td><td>Cheese</td><td>Lys</td><td>Leu</td><td>Thr</td><td>Val</td><td>Asp</td><td>Lys</td><td>Cheese</td>

<td></td><td></td><td></td><td> 420</td><td></td><td></td><td></td><td></td><td> 425</td><td></td><td></td><td></td><td></td><td> 430</td><td></td><td></td>

<td>Arg</td><td>Trp</td><td>Gin</td><td>Gin</td><td>Gly</td><td>Asn</td><td>Val</td><td>Phe</td><td>Cheese</td><td>Cys</td><td>Cheese</td><td>Val</td><td>Underworld</td><td>His</td><td>Glu</td><td>Ala</td>

<td></td><td></td><td> 435</td><td></td><td></td><td></td><td></td><td> 440</td><td></td><td></td><td></td><td></td><td> 445</td><td></td><td></td><td></td>

<td>Leu</td><td>His</td><td>Asn</td><td>His</td><td>Tyr</td><td>Thr</td><td>Gin</td><td>Lys</td><td>Cheese</td><td>Leu</td><td>Cheese</td><td>Leu</td><td>Cheese</td><td>Pro</td><td>Gly</td><td>Bald</td>

450 455 460 &lt;210&gt; 3 &lt;211&gt; 163 &lt;212&gt; PRT &lt;213&gt; Homo sapiens &lt;400&gt; 3

<td colspan="2">Pro Gly 1</td><td>Arg</td><td>Cheese</td><td colspan="2">Leu Arg 5</td><td>Leu</td><td>Cheese</td><td>Cys</td><td colspan="2">Ala Ala 10</td><td>Cheese</td><td>Gly</td><td>Phe</td><td>Thr 15</td><td>Phe</td>

<td>Cheese</td><td>Cheese</td><td>His</td><td>Gly</td><td>How much</td><td>His</td><td>Trp</td><td>Val</td><td>Arg</td><td>Gin</td><td>Ala</td><td>Pro</td><td>Gly</td><td>Lys</td><td>Gly</td><td>Leu</td>

<td></td><td></td><td></td><td> 20</td><td></td><td></td><td></td><td></td><td> 25</td><td></td><td></td><td></td><td></td><td> 30</td><td></td><td></td>

<td>Glu</td><td>Trp</td><td>Val</td><td>Ala</td><td>Val</td><td>How much</td><td>Trp</td><td>Tyr</td><td>Asp</td><td>Gly</td><td>Arg</td><td>Asn</td><td>Lys</td><td>Asp</td><td>Tyr</td><td>Ala</td>

<td></td><td></td><td> 35</td><td></td><td></td><td></td><td></td><td> 40</td><td></td><td></td><td></td><td></td><td> 45</td><td></td><td></td><td></td>

<td>Asp</td><td>Cheese</td><td>Val</td><td>Lys</td><td>Gly</td><td>Arg</td><td>Phe</td><td>Thr</td><td>How much</td><td>Cheese</td><td>Arg</td><td>Asp</td><td>Asn</td><td>Cheese</td><td>Lys</td><td>Lys</td>

<td></td><td> 50</td><td></td><td></td><td></td><td></td><td> 55</td><td></td><td></td><td></td><td></td><td> 60</td><td></td><td></td><td></td><td></td>

<td>Thr</td><td>Leu</td><td>Tyr</td><td>Leu</td><td>Gin</td><td>Underworld</td><td>Asn</td><td>Cheese</td><td>Leu</td><td>Arg</td><td>Ala</td><td>Glu</td><td>Asp</td><td>Thr</td><td>Ala</td><td>Val</td>

<td> 65</td><td></td><td></td><td></td><td></td><td> 70</td><td></td><td></td><td></td><td></td><td> 75</td><td></td><td></td><td></td><td></td><td> 80</td>

Tyr Tyr Cys Ala Arg V-.il Ala Pro Leu. Gly Pro Leu Asp Tyr Trp Gly

90 95

PL 214 003 Β1

<td>Gin</td><td>dy</td><td>Thr</td><td>Leu 100</td><td>Val</td><td>Thr</td><td>Val</td><td>Cheese</td><td>Cheese 105</td><td>Ala</td><td>Cheese</td><td>Thr</td><td>Lys</td><td>Gly 110</td><td>Pro</td><td>Cheese</td>

<td>Val</td><td>Phe</td><td>Pro</td><td>Leu</td><td>Ala</td><td>Pro</td><td>Cys</td><td>Cheese</td><td>Arg</td><td>Cheese</td><td>Thr</td><td>Cheese</td><td>Glu</td><td>Cheese</td><td>Thr</td><td>Ala</td>

<td></td><td></td><td> 115</td><td></td><td></td><td></td><td></td><td> 120</td><td></td><td></td><td></td><td></td><td> 125</td><td></td><td></td><td></td>

<td>Ala</td><td>Leu</td><td>Gly</td><td>Cys</td><td>Leu</td><td>Val</td><td>Lys</td><td>Asp</td><td>Tyr</td><td>Phe</td><td>Pro</td><td>Glu</td><td>Pro</td><td>Val</td><td>Thr</td><td>Val</td>

<td></td><td> 130</td><td></td><td></td><td></td><td></td><td> 135</td><td></td><td></td><td></td><td></td><td> 140</td><td></td><td></td><td></td><td></td>

<td>Cheese</td><td>Trp</td><td>Asn</td><td>Cheese</td><td>Gly</td><td>Ala</td><td>Leu</td><td>Thr</td><td>Cheese</td><td>Gly</td><td>Val</td><td>His</td><td>Thr</td><td>Phe</td><td>Pro</td><td>Ala</td>

<td> 145</td><td></td><td></td><td></td><td></td><td> 150</td><td></td><td></td><td></td><td></td><td> 155</td><td></td><td></td><td></td><td></td><td> 160</td>

<td>Val</td><td>Leu</td><td>Gin</td><td></td><td></td><td></td><td></td><td></td><td></td><td></td><td></td><td></td><td></td><td></td><td></td><td></td>

&lt;210&gt; 4 &lt;211&gt; 463 &lt;212&gt; PRT &lt;213&gt; Homo sapiens

<td colspan="8"> &lt;400&gt; 4</td><td rowspan="2">Phe</td><td rowspan="2">Leu 10</td><td rowspan="2">Val</td><td rowspan="2">Ala</td><td rowspan="2">Leu</td><td rowspan="2">Leu</td><td rowspan="2">Arg 15</td><td rowspan="2">Gly</td>

<td>Underworld 1</td><td>Glu</td><td colspan="2">Phe Gly</td><td>Leu 5</td><td>Cheese</td><td>Trp</td><td>Val</td>

<td>Val</td><td>Gin</td><td>Cys</td><td>Gin</td><td>Val</td><td>Gin</td><td>Leu</td><td>Val</td><td>Glu</td><td>Cheese</td><td>Gly</td><td>Gly</td><td>Gly</td><td>Val</td><td>Val</td><td>Glu</td>

<td></td><td></td><td></td><td> 20</td><td></td><td></td><td></td><td></td><td> 25</td><td></td><td></td><td></td><td></td><td> 30</td><td></td><td></td>

<td>Pro</td><td>Gly</td><td>Arg</td><td>Cheese</td><td>Leu</td><td>Arg</td><td>Leu</td><td>Cheese</td><td>Cys</td><td>Thr</td><td>Ala</td><td>Cheese</td><td>Gly</td><td>Phe</td><td>Thr</td><td>Phe</td>

<td></td><td></td><td> 35</td><td></td><td></td><td></td><td></td><td> 40</td><td></td><td></td><td></td><td></td><td> 45</td><td></td><td></td><td></td>

<td>Cheese</td><td>Cheese</td><td>Tyr</td><td>Gly</td><td>Underworld</td><td>His</td><td>Trp</td><td>Val</td><td>Arg</td><td>Gin</td><td>Ala</td><td>Pro</td><td>Gly</td><td>Lys</td><td>dy</td><td>Leu</td>

<td></td><td> 50</td><td></td><td></td><td></td><td></td><td> 55</td><td></td><td></td><td></td><td></td><td> 60</td><td></td><td></td><td></td><td></td>

<td>Glu</td><td>Trp</td><td>Val</td><td>Ala</td><td>val</td><td>How much</td><td>Trp</td><td>Tyr</td><td>Asp</td><td>Gly</td><td>Cheese</td><td>Asn</td><td>Lys</td><td>His</td><td>Tyr</td><td>Ala</td>

<td> 65</td><td></td><td></td><td></td><td></td><td> 70</td><td></td><td></td><td></td><td></td><td> 75</td><td></td><td></td><td></td><td></td><td> 80</td>

<td>Asp</td><td>Cheese</td><td>Ala</td><td>Lys</td><td>Gly</td><td>Arg</td><td>Phe</td><td>Thr</td><td>How much</td><td>Cheese</td><td>Arg</td><td>Asp</td><td>Asn</td><td>Cheese</td><td>Lys</td><td>Asn</td>

<td></td><td></td><td></td><td></td><td> 85</td><td></td><td></td><td></td><td></td><td> 90</td><td></td><td></td><td></td><td></td><td> 95</td><td></td>

<td>Thr</td><td>Leu</td><td>Tyr</td><td>Leu</td><td>Gin</td><td>Underworld</td><td>Asn</td><td>Cheese</td><td>Leu</td><td>Arg</td><td>Al a</td><td>Glu</td><td>Asp</td><td>Thr</td><td>Ala</td><td>Val</td>

<td></td><td></td><td></td><td> 100</td><td></td><td></td><td></td><td></td><td> 105</td><td></td><td></td><td></td><td></td><td> 110</td><td></td><td></td>

<td>Tyr</td><td>Tyr</td><td>Cys</td><td>Ala</td><td>Arg</td><td>Ala</td><td>θίγ</td><td>Leu</td><td>leu</td><td>Gly</td><td>Tyr</td><td>Phe</td><td>Asp</td><td>Tyr</td><td>Trp</td><td>Gly</td>

<td></td><td></td><td> 115</td><td></td><td></td><td></td><td></td><td> 120</td><td></td><td></td><td></td><td></td><td> 125</td><td></td><td></td><td></td>

<td>Gin</td><td>Gly</td><td>Thr</td><td>Leu</td><td>Val</td><td>Thr</td><td>Val</td><td>Cheese</td><td>Cheese</td><td>Ala</td><td>Cheese</td><td>Thr</td><td>Lys</td><td>dy</td><td>Pro</td><td>Cheese</td>

<td></td><td> 130</td><td></td><td></td><td></td><td></td><td> 135</td><td></td><td></td><td></td><td></td><td> 140</td><td></td><td></td><td></td><td></td>

<td>Val</td><td>Phe</td><td>Pro</td><td>Leu</td><td>Ala</td><td>Pro</td><td>Cys</td><td>Cheese</td><td>Arg</td><td>Cheese</td><td>Thr</td><td>Cheese</td><td>Glu</td><td>Cheese</td><td>Thr</td><td>Ala</td>

<td> 145</td><td></td><td></td><td></td><td></td><td> 150</td><td></td><td></td><td></td><td></td><td> 155</td><td></td><td></td><td></td><td></td><td> 160</td>

<td>Ala</td><td>Leu</td><td>Gly</td><td>Cys</td><td>Leu</td><td>Vai</td><td>Lys</td><td>Asp</td><td>Tyr</td><td>Phe</td><td>Pro</td><td>Glu</td><td>Pro</td><td>Val</td><td>Thr</td><td>Val</td>

<td></td><td></td><td></td><td></td><td> 165</td><td></td><td></td><td></td><td></td><td> 170</td><td></td><td></td><td></td><td></td><td> 175</td><td></td>

<td>Cheese</td><td>Trp</td><td>Asn</td><td>Cheese</td><td>Gly</td><td>Ala</td><td>L. ^,eu</td><td>Thr</td><td>Cheese</td><td>dy</td><td>Val</td><td>His</td><td>Thr</td><td>Phe</td><td>Pro</td><td>Ala</td>

<td></td><td></td><td></td><td> 180</td><td></td><td></td><td></td><td></td><td> 185</td><td></td><td></td><td></td><td></td><td> 190</td><td></td><td></td>

<td>Val</td><td>Leu</td><td>Gin</td><td>Cheese</td><td>Cheese</td><td>Gly</td><td></td><td>Tyr</td><td>Cheese</td><td>Leu</td><td>Cheese</td><td>Cheese</td><td>val</td><td>val</td><td>Thr</td><td>Val</td>

PL 214 003 Β1

<td></td><td colspan="4"> 195</td><td colspan="6"> 200</td><td colspan="5"> 205</td>

<td>Pro</td><td>Cheese</td><td>Cheese</td><td>Asn</td><td>Phe</td><td>Gly</td><td>Thr</td><td>Gin</td><td>Thr</td><td>Tyr</td><td>Thr</td><td>Cys</td><td>Asn</td><td>Yal</td><td>Asp</td><td>His</td>

<td></td><td>2x0</td><td></td><td></td><td></td><td></td><td> 215</td><td></td><td></td><td></td><td></td><td> 220</td><td></td><td></td><td></td><td></td>

<td>Lys</td><td>Pro</td><td>Cheese</td><td>Asn</td><td>Thr</td><td>Lys</td><td>Val</td><td>Asp</td><td>Lys</td><td>Thr</td><td>Val</td><td>Glu</td><td>Arg</td><td>Lys</td><td>Cys</td><td>cys</td>

<td> 225</td><td></td><td></td><td></td><td></td><td> 230</td><td></td><td></td><td></td><td></td><td> 235</td><td></td><td></td><td></td><td></td><td> 240</td>

<td>Val</td><td>Glu</td><td>Cys</td><td>Pro</td><td>Pro</td><td>Cys</td><td>Pro</td><td>Ala</td><td>Pro</td><td>Pro</td><td>Val</td><td>Ala</td><td>Gly</td><td>Pro</td><td>Cheese</td><td>Val</td>

<td></td><td></td><td></td><td></td><td> 245</td><td></td><td></td><td></td><td></td><td> 250</td><td></td><td></td><td></td><td></td><td> 255</td><td></td>

<td>Phe</td><td>Leu</td><td>Phe</td><td>Pro</td><td>Pro</td><td>Lys</td><td>Pro</td><td>Lys</td><td>Asp</td><td>Thr</td><td>Leu</td><td>Underworld</td><td>How much</td><td>Cheese</td><td>Arg</td><td>Thr</td>

<td></td><td></td><td></td><td> 260</td><td></td><td></td><td></td><td></td><td> 265</td><td></td><td></td><td></td><td></td><td> 270</td><td></td><td></td>

<td>Pro</td><td>Glu</td><td>Val</td><td>Thr</td><td>Cys</td><td>Val</td><td>Val</td><td>Val</td><td>Asp</td><td>Val</td><td>Cheese</td><td>His</td><td>Glu</td><td>Asp</td><td>Pro</td><td>Glu</td>

<td></td><td></td><td> 275</td><td></td><td></td><td></td><td></td><td> 280</td><td></td><td></td><td></td><td></td><td> 285</td><td></td><td></td><td></td>

<td>Val</td><td>Gin</td><td>Phe</td><td>Asn</td><td>Trp</td><td>Tyr</td><td>Val</td><td>Asp</td><td>Gly</td><td>Val</td><td>Glu</td><td>Val</td><td>His</td><td>Asn</td><td>Ala</td><td>Lys</td>

<td></td><td> 290</td><td></td><td></td><td></td><td></td><td> 295</td><td></td><td></td><td></td><td></td><td> 300</td><td></td><td></td><td></td><td></td>

<td>Thr</td><td>Lys</td><td>Pro</td><td>Arg</td><td>Glu</td><td>Glu</td><td>Gin</td><td>Phe</td><td>Asn</td><td>Cheese</td><td>Thr</td><td>Phe</td><td>Arg</td><td>Val</td><td>Val</td><td>Cheese</td>

<td> 305</td><td></td><td></td><td></td><td></td><td> 310</td><td></td><td></td><td></td><td></td><td> 315</td><td></td><td></td><td></td><td></td><td> 320</td>

<td>Val</td><td>Leu</td><td>Thr</td><td>Val</td><td>Val</td><td>His</td><td>Gin</td><td>Asp</td><td>Trp</td><td>Leu</td><td>Asn</td><td>Gly</td><td>Lys</td><td>Glu</td><td>Tyr</td><td>Lys</td>

<td></td><td></td><td></td><td></td><td> 325</td><td></td><td></td><td></td><td></td><td> 330</td><td></td><td></td><td></td><td></td><td> 335</td><td></td>

<td>Cys</td><td>Lys</td><td>Val</td><td>Cheese</td><td>Asn</td><td>Lys</td><td>dy</td><td>Leu</td><td>Pro</td><td>Ala</td><td>Pro</td><td>How much</td><td>Glu</td><td>Lys</td><td>Thr</td><td>How much</td>

<td></td><td></td><td></td><td> 340</td><td></td><td></td><td></td><td></td><td> 345</td><td></td><td></td><td></td><td></td><td> 350</td><td></td><td></td>

<td>Cheese</td><td>Lys</td><td>Thr</td><td>Lys</td><td>Gly</td><td>Gin</td><td>Pro</td><td>Arg</td><td>Glu</td><td>Pro</td><td>Gin</td><td>Val</td><td>Tyr</td><td>Thr</td><td>Leu</td><td>Pro</td>

<td></td><td></td><td> 355</td><td></td><td></td><td></td><td></td><td> 360</td><td></td><td></td><td></td><td></td><td> 365</td><td></td><td></td><td></td>

<td>Pro</td><td>Cheese</td><td>Arg</td><td>Glu</td><td>Glu</td><td>Underworld</td><td>Thr</td><td>Lys</td><td>Asn</td><td>Gin</td><td>Val</td><td>Cheese</td><td>Leu</td><td>Thr</td><td>Cys</td><td>Leu</td>

<td></td><td> 370</td><td></td><td></td><td></td><td></td><td> 375</td><td></td><td></td><td></td><td></td><td> 380</td><td></td><td></td><td></td><td></td>

<td>Val</td><td>Lys</td><td>Gly</td><td>Phe</td><td>Tyr</td><td>Pro</td><td>Cheese</td><td>Asp</td><td>How much</td><td>Ala</td><td>Val</td><td>Glu</td><td>Trp</td><td>Glu</td><td>Cheese</td><td>Asn</td>

<td> 385</td><td></td><td></td><td></td><td></td><td> 390</td><td></td><td></td><td></td><td></td><td> 395</td><td></td><td></td><td></td><td></td><td> 4 00</td>

<td>Gly</td><td>Gin</td><td>Pro</td><td>Glu</td><td>Asn</td><td>Asn</td><td>Tyr</td><td>Lys</td><td>Thr</td><td>Thr</td><td>Pro</td><td>Pro</td><td>Underworld</td><td>Leu</td><td>Asp</td><td>Cheese</td>

<td></td><td></td><td></td><td></td><td> 405</td><td></td><td></td><td></td><td></td><td> 410</td><td></td><td></td><td></td><td></td><td> 415</td><td></td>

<td>Asp</td><td>Gly</td><td>Cheese</td><td>Phe</td><td>Phe</td><td>Leu</td><td>Tyr</td><td>Cheese</td><td>Lys</td><td>Leu</td><td>Thr</td><td>Val</td><td>Asp</td><td>Lys</td><td>Cheese</td><td>Arg</td>

<td></td><td></td><td></td><td> 420</td><td></td><td></td><td></td><td></td><td> 425</td><td></td><td></td><td></td><td></td><td> 430</td><td></td><td></td>

<td>Trp</td><td>Gin</td><td>Gin</td><td>Gly</td><td>Asn</td><td>Val</td><td>Phe</td><td>Cheese</td><td>Cys</td><td>Cheese</td><td>Val</td><td>Underworld</td><td>Hi s</td><td>Glu</td><td>Al a</td><td>Leu</td>

<td></td><td></td><td> 435</td><td></td><td></td><td></td><td></td><td> 440</td><td></td><td></td><td></td><td></td><td> 445</td><td></td><td></td><td></td>

<td>His</td><td>Asn</td><td>His</td><td>Tyr</td><td>Thr</td><td>Gin</td><td>Lys</td><td>Cheese</td><td>Leu</td><td>Cheese</td><td>Leu</td><td>Cheese</td><td>Pro</td><td>Gly</td><td>Lys</td><td></td>

<td></td><td> 450</td><td></td><td></td><td></td><td></td><td> 455</td><td></td><td></td><td></td><td></td><td> 4 60</td><td></td><td></td><td></td><td></td>

&lt;210&gt; 5 &lt;211&gt; 169 &lt;212&gt; PRT &lt;213&gt; Homo sapiens &lt;400&gt; 5

Gly Val Val Gin Pro Gly Arg Ser Leu Arg Leu Ser Cys Ala Ala Ser

PL 214 003 Β1

<td colspan="3"> 1</td><td colspan="2"> 5</td><td colspan="8"> 10</td><td colspan="3"> 15</td>

<td>Giy</td><td>Phe</td><td>Thr</td><td>Phe</td><td>Cheese</td><td>Cheese</td><td>Tyr</td><td>Gly</td><td>Underworld</td><td>His</td><td>Trp</td><td>Va 1</td><td>Arg</td><td>Gin</td><td>Ala</td><td>Pro</td>

<td></td><td></td><td></td><td> 20</td><td></td><td></td><td></td><td></td><td> 25</td><td></td><td></td><td></td><td></td><td> 3 0</td><td></td><td></td>

<td>Gly</td><td>Lys</td><td>Gly</td><td>Leu</td><td>Glu</td><td>Trp</td><td>Val</td><td>Ala</td><td>Val</td><td>Background</td><td>Trp</td><td>Tyr</td><td>Asp</td><td>Gly</td><td>Cheese</td><td>Asn</td>

<td></td><td></td><td> 35</td><td></td><td></td><td></td><td></td><td> 40</td><td></td><td></td><td></td><td></td><td> 45</td><td></td><td></td><td></td>

<td>Lys</td><td>Tyr</td><td>Tyr</td><td>Ala</td><td>Asp</td><td>Cheese</td><td>Val</td><td>Lys</td><td>Gly</td><td>Arg</td><td>Phe</td><td>Thr</td><td>How much</td><td>Cheese</td><td>Arg</td><td>Asp</td>

<td></td><td> 50</td><td></td><td></td><td></td><td></td><td> 55</td><td></td><td></td><td></td><td></td><td> 60</td><td></td><td></td><td></td><td></td>

<td>Asn</td><td>Cheese</td><td>Lys</td><td>Asn</td><td>Thr</td><td>Leu</td><td>Tyr</td><td>Leu</td><td>Gin</td><td>Underworld</td><td>Asn</td><td>Cheese</td><td>Leu</td><td>Arg</td><td>Ala</td><td>Glu</td>

<td> 65</td><td></td><td></td><td></td><td></td><td> 70</td><td></td><td></td><td></td><td></td><td> 75</td><td></td><td></td><td></td><td></td><td> 80</td>

<td>Asp</td><td>Thr</td><td>Ala</td><td>Val</td><td>Tyr</td><td>Tyr</td><td>Cys</td><td>Ala</td><td>Arg</td><td>Gly</td><td>Ala</td><td>Arg</td><td>How much</td><td>How much</td><td>Thr</td><td>Pro</td>

<td></td><td></td><td></td><td></td><td> 85</td><td></td><td></td><td></td><td></td><td> 90</td><td></td><td></td><td></td><td></td><td> 95</td><td></td>

<td>Cys</td><td>Underworld</td><td>Asp</td><td>Val</td><td>Trp</td><td>Gly</td><td>Gin</td><td>Gly</td><td>Thr</td><td>Thr</td><td>Val</td><td>Thr</td><td>Val</td><td>Cheese</td><td>Cheese</td><td>Ala</td>

<td></td><td></td><td></td><td> 100</td><td></td><td></td><td></td><td></td><td> 105</td><td></td><td></td><td></td><td></td><td> 110</td><td></td><td></td>

<td>Cheese</td><td>Thr</td><td>Lys</td><td>Gly</td><td>Pro</td><td>Cheese</td><td>val</td><td>Phe</td><td>Pro</td><td>Leu</td><td>Ala</td><td>Pro</td><td>cys</td><td>Cheese</td><td>Arg</td><td>Cheese</td>

<td></td><td></td><td> 115</td><td></td><td></td><td></td><td></td><td> 120</td><td></td><td></td><td></td><td></td><td> 125</td><td></td><td></td><td></td>

<td>Thr</td><td>Cheese</td><td>Glu</td><td>Cheese</td><td>Thr</td><td>Ala</td><td>Ala</td><td>Leu</td><td>Gly</td><td>Cys</td><td>Leu</td><td>val</td><td>Lys</td><td>Asp</td><td>Tyr</td><td>Phe</td>

<td></td><td> 130</td><td></td><td></td><td></td><td></td><td> 135</td><td></td><td></td><td></td><td></td><td> 140</td><td></td><td></td><td></td><td></td>

<td>Pro</td><td>Glu</td><td>Pro</td><td>Val</td><td>Thr</td><td>Val</td><td>Cheese</td><td>Trp</td><td>Asn</td><td>Cheese</td><td>Gly</td><td>Ar a</td><td>Leu</td><td>Thr</td><td>Cheese</td><td>Gly</td>

<td> 145</td><td></td><td></td><td></td><td></td><td> 150</td><td></td><td></td><td></td><td></td><td> 155</td><td></td><td></td><td></td><td></td><td> 160</td>

<td>Val</td><td>His</td><td>Thr</td><td>Phe</td><td>Pro</td><td>Ala</td><td>Val</td><td>Leu</td><td>Gin</td><td></td><td></td><td></td><td></td><td></td><td></td><td></td>

165 &lt;210&gt; 6 &lt;211&gt; 167 &lt;212&gt; PRT &lt;213&gt; Homo sapiens &lt;400&gt; 6

<td>Gly 1</td><td>Val</td><td colspan="2">Val Gin</td><td>Pro 5</td><td>Gly</td><td>Arg</td><td>Cheese</td><td>Leu</td><td>Arg 10</td><td>Leu</td><td colspan="2">Cheese Cys</td><td>Val</td><td>Ala 15</td><td>Cheese</td>

<td>Gly</td><td>Phe</td><td>How much</td><td>Phe</td><td>Cheese</td><td>Cheese</td><td>His</td><td>Gly</td><td>How much</td><td>His</td><td>Trp</td><td>Val</td><td>Arg</td><td>Gin</td><td>Ala</td><td>Pro</td>

<td></td><td></td><td></td><td> 20</td><td></td><td></td><td></td><td></td><td> 25</td><td></td><td></td><td></td><td></td><td> 30</td><td></td><td></td>

<td>Gly</td><td>Lys</td><td>Gly</td><td>Leu</td><td>Glu</td><td>Trp</td><td>Val</td><td>Ala</td><td>Val</td><td>How much</td><td>Trp</td><td>Tyr</td><td>Asp</td><td>dy</td><td>Arg</td><td>Asn</td>

<td></td><td></td><td> 35</td><td></td><td></td><td></td><td></td><td> 40</td><td></td><td></td><td></td><td></td><td> 45</td><td></td><td></td><td></td>

<td>Lys</td><td>Asp</td><td>Tyr</td><td>Ala</td><td>Asp</td><td>Cheese</td><td>Val</td><td>Lys</td><td>Gly</td><td>Arg</td><td>Phe</td><td>Thr</td><td>How much</td><td>Cheese</td><td>Arg</td><td>Asp</td>

<td></td><td> 50</td><td></td><td></td><td></td><td></td><td> 55</td><td></td><td></td><td></td><td></td><td> 60</td><td></td><td></td><td></td><td></td>

<td>Asn</td><td>Cheese</td><td>Lys</td><td>Asn</td><td>Thr</td><td>Leu</td><td>Tyr</td><td>Leu</td><td>Gin</td><td>Underworld</td><td>Asn</td><td>Cheese</td><td>Leu</td><td>Ara</td><td>Ala</td><td>Glu</td>

<td> 65</td><td></td><td></td><td></td><td></td><td> 70</td><td></td><td></td><td></td><td></td><td> 75</td><td></td><td></td><td></td><td></td><td> 80</td>

<td>Asp</td><td>Thr</td><td>Ala</td><td>val</td><td>Tyr</td><td>Tyr</td><td>Cys</td><td>Ala</td><td>Arg</td><td>Val</td><td>Ala</td><td>Pro</td><td>Leu</td><td>Gly</td><td>Pro</td><td>Leu</td>

<td></td><td></td><td></td><td></td><td> 85</td><td></td><td></td><td></td><td></td><td> 90</td><td></td><td></td><td></td><td></td><td> 95</td><td></td>

Asp Tyr Trp Gly Gin Gly Thr Leu Val Thr Val Ser Ser Ala Ser Thr

PL 214 003 Β1

<td></td><td></td><td></td><td> 100</td><td></td><td></td><td></td><td></td><td> 105</td><td></td><td></td><td></td><td></td><td> 110</td><td></td><td></td>

<td>Lys</td><td>Gly</td><td>Pro</td><td>Cheese</td><td>Val</td><td>Phe</td><td>Pro</td><td>Leu</td><td>Ala</td><td>Pro</td><td>cys</td><td>Cheese</td><td>Arg</td><td>Cheese</td><td>Thr</td><td>Cheese</td>

<td></td><td></td><td> 115</td><td></td><td></td><td></td><td></td><td> 120</td><td></td><td></td><td></td><td></td><td> 125</td><td></td><td></td><td></td>

<td>Glu</td><td>Cheese</td><td>Thr</td><td>Al a</td><td>Ala</td><td>Leu</td><td>Gly</td><td>Cys</td><td>Leu</td><td>Val</td><td>Lys</td><td>Asp</td><td>Tyr</td><td>Phe</td><td>Pro</td><td>Glu</td>

<td></td><td> 130</td><td></td><td></td><td></td><td></td><td> 135</td><td></td><td></td><td></td><td></td><td> 140</td><td></td><td></td><td></td><td></td>

<td>Pro</td><td>Val</td><td>Thr</td><td>Val</td><td>Cheese</td><td>Trp</td><td>Asn</td><td>Cheese</td><td>Gly</td><td>Ala</td><td>Leu</td><td>Thr</td><td>Cheese</td><td>Gly</td><td>Val</td><td>His</td>

<td> 145</td><td></td><td></td><td></td><td></td><td> 150</td><td></td><td></td><td></td><td></td><td> 155</td><td></td><td></td><td></td><td></td><td> 160</td>

<td>Thr</td><td>Phe</td><td>Pro</td><td>Ala</td><td>Val</td><td>Leu</td><td>Gin</td><td></td><td></td><td></td><td></td><td></td><td></td><td></td><td></td><td></td>

165 &lt;210&gt; Ί &lt;211&gt; 172 &lt;212&gt; PRT

<td colspan="3">&lt;213&gt; Homo</td><td colspan="13">sapiens</td>

<td colspan="2"> &lt;400&gt; 7</td><td></td><td></td><td></td><td></td><td></td><td></td><td></td><td></td><td></td><td></td><td></td><td></td><td></td><td></td>

<td>Cheese</td><td>Gly</td><td>Pro</td><td>Gly</td><td>Leu</td><td>Val</td><td>Lys</td><td>Pro</td><td>Cheese</td><td>Gin</td><td>Background</td><td>Leu</td><td>Cheese</td><td>Leu</td><td>Thr</td><td>cys</td>

<td> 1</td><td></td><td></td><td></td><td> 5</td><td></td><td></td><td></td><td></td><td> 10</td><td></td><td></td><td></td><td></td><td> 15</td><td></td>

<td>Thr</td><td>Val</td><td>Cheese</td><td>Gly</td><td>Gly</td><td>Cheese</td><td>How much</td><td>Cheese</td><td>Cheese</td><td>Gly</td><td>Gly</td><td>His</td><td>Tyr</td><td>Trp</td><td>Cheese</td><td>Trp</td>

<td></td><td></td><td></td><td> 20</td><td></td><td></td><td></td><td></td><td> 25</td><td></td><td></td><td></td><td></td><td> 30</td><td></td><td></td>

<td>how much</td><td>Arg</td><td>Gin</td><td>His</td><td>Pro</td><td>Gly</td><td>Lys</td><td>Gly</td><td>Leu</td><td>Glu</td><td>Trp</td><td>How much</td><td>Gly</td><td>Tyr</td><td>Background</td><td>Tyr</td>

<td></td><td></td><td> 35</td><td></td><td></td><td></td><td></td><td> 40</td><td></td><td></td><td></td><td></td><td> 45</td><td></td><td></td><td></td>

<td>Tyr</td><td>how much</td><td>Gly</td><td>Asn</td><td>Thr</td><td>Tyr</td><td>Tyr</td><td>Asn</td><td>Pro</td><td>Cheese</td><td>Leu</td><td>Lys</td><td>Cheese</td><td>Arg</td><td>Val</td><td>Thr</td>

<td></td><td> 50</td><td></td><td></td><td></td><td></td><td> 55</td><td></td><td></td><td></td><td></td><td> 60</td><td></td><td></td><td></td><td></td>

<td>How much</td><td>Cheese</td><td>val</td><td>Asp</td><td>Thr</td><td>Cheese</td><td>Lys</td><td>Asn</td><td>Gin</td><td>Phe</td><td>Cheese</td><td>Leu</td><td>Lys</td><td>Leu</td><td>Cheese</td><td>Cheese</td>

<td> 65</td><td></td><td></td><td></td><td></td><td> 70</td><td></td><td></td><td></td><td></td><td> 75</td><td></td><td></td><td></td><td></td><td> 80</td>

<td>Val</td><td>Thr</td><td>Ala</td><td>Ala</td><td>Asp</td><td>Thr</td><td>Ala</td><td>Val</td><td>Tyr</td><td>Tyr</td><td>Cys</td><td>Ala</td><td>Arg</td><td>Asp</td><td>Cheese</td><td>Gly</td>

<td></td><td></td><td></td><td></td><td> 85</td><td></td><td></td><td></td><td></td><td> 90</td><td></td><td></td><td></td><td></td><td> 95</td><td></td>

<td>Asp</td><td>Tyr</td><td>Tyr</td><td>Gly</td><td>How much</td><td>Asp</td><td>Val</td><td>Trp</td><td>Gly</td><td>Gin</td><td>Gly</td><td>Thr</td><td>Thr</td><td>vai</td><td>Thr</td><td>Val</td>

<td></td><td></td><td></td><td> 100</td><td></td><td></td><td></td><td></td><td> 105</td><td></td><td></td><td></td><td></td><td> 110</td><td></td><td></td>

<td>Cheese</td><td>Cheese</td><td>Ala</td><td>Cheese</td><td>Thr</td><td>Lys</td><td>Gly</td><td>Pro</td><td>Cheese</td><td>Val</td><td>Phe</td><td>Pro</td><td>Leu</td><td>Al a</td><td>Pro</td><td>Cys</td>

<td></td><td></td><td> 115</td><td></td><td></td><td></td><td></td><td> 120</td><td></td><td></td><td></td><td></td><td> 125</td><td></td><td></td><td></td>

<td>Cheese</td><td>Arg</td><td>Cheese</td><td>Thr</td><td>Cheese</td><td>Glu</td><td>Cheese</td><td>Thr</td><td>Ala</td><td>Ala</td><td>Leu</td><td>Gly</td><td>Cys</td><td>Leu</td><td>Val</td><td>Lys</td>

<td></td><td> 130</td><td></td><td></td><td></td><td></td><td> 135</td><td></td><td></td><td></td><td></td><td> 140</td><td></td><td></td><td></td><td></td>

<td>Asp</td><td>Tyr</td><td>Phe</td><td>Pro</td><td>Glu</td><td>Pro</td><td>Val</td><td>Thr</td><td>Val</td><td>Cheese</td><td>Trp</td><td>Asn</td><td>Cheese</td><td>Gly</td><td>Ala</td><td>Leu</td>

<td> 145</td><td></td><td></td><td></td><td></td><td> 150</td><td></td><td></td><td></td><td></td><td> 155</td><td></td><td></td><td></td><td></td><td> 160</td>

<td>Thr</td><td>Cheese</td><td>Gly</td><td>Val</td><td>His</td><td>Thr</td><td>Phe</td><td>Pro</td><td>Ala</td><td>vai</td><td>Leu</td><td>Gin</td><td></td><td></td><td></td><td></td>

165 170 &lt;210&gt; 8 &lt;211&gt; 153 &lt;212&gt; PRT

PL 214 003 Β1 &lt;213&gt; Homo sapiens &lt;400&gt; 8

<td>Pro 1</td><td colspan="2">Gly Arg</td><td>Cheese</td><td>Leu 5</td><td>Arg</td><td>Leu</td><td>Cheese</td><td colspan="2">Cys Ala 10</td><td>Ala</td><td>Cheese</td><td>Gly</td><td>Phe</td><td>Thr 15</td><td>Phe</td>

<td>Cheese</td><td>Cheese</td><td>His</td><td>Gly</td><td>How much</td><td>His</td><td>Trp</td><td>Val</td><td>Arg</td><td>Gin</td><td>Ala</td><td>Pro</td><td>Gly</td><td>Lys</td><td>Gly</td><td>Leu</td>

<td></td><td></td><td></td><td> 20</td><td></td><td></td><td></td><td></td><td> 25</td><td></td><td></td><td></td><td></td><td> 30</td><td></td><td></td>

<td>Glu</td><td>Trp</td><td>Val</td><td>Ala</td><td>Val</td><td>How much</td><td>Trp</td><td>Tyr</td><td>Asp</td><td>Gly</td><td>Arg</td><td>Asn</td><td>Lys</td><td>Asp</td><td>Tyr</td><td>Ala</td>

<td></td><td></td><td> 35</td><td></td><td></td><td></td><td></td><td> 40</td><td></td><td></td><td></td><td></td><td> 45</td><td></td><td></td><td></td>

<td>Asp</td><td>Cheese</td><td>Val</td><td>Lys</td><td>Gly</td><td>Arg</td><td>Phe</td><td>Thr</td><td>How much</td><td>Cheese</td><td>Arg</td><td>Asp</td><td>Asn</td><td>Cheese</td><td>Lys</td><td>Asn</td>

<td></td><td> 50</td><td></td><td></td><td></td><td></td><td> 55</td><td></td><td></td><td></td><td></td><td> 60</td><td></td><td></td><td></td><td></td>

<td>Thr</td><td>Leu</td><td>Tyr</td><td>Leu</td><td>Gin</td><td>Underworld</td><td>Asn</td><td>Cheese</td><td>Leu</td><td>Arg</td><td>Ala</td><td>Glu</td><td>Asp</td><td>Thr</td><td>Ala</td><td>Val</td>

<td> 65</td><td></td><td></td><td></td><td></td><td>7G</td><td></td><td></td><td></td><td></td><td> 75</td><td></td><td></td><td></td><td></td><td> 80</td>

<td>Tyr</td><td>Tyr</td><td>Cys</td><td>Ala</td><td>Arg</td><td>Val</td><td>Ala</td><td>Pro</td><td>Leu</td><td>Gly</td><td>Pro</td><td>Leu</td><td>Asp</td><td>Φ yy</td><td>Trp</td><td>Gly</td>

<td></td><td></td><td></td><td></td><td> 85</td><td></td><td></td><td></td><td></td><td> 90</td><td></td><td></td><td></td><td></td><td> 95</td><td></td>

<td>Gin</td><td>Gly</td><td>Thr</td><td>Leu</td><td>val</td><td>Thr</td><td>Val</td><td>Cheese</td><td>Cheese</td><td>Ala</td><td>Cheese</td><td>Thr</td><td>Lys</td><td>Gly</td><td>Pro</td><td>Cheese</td>

<td></td><td></td><td></td><td> 100</td><td></td><td></td><td></td><td></td><td> 105</td><td></td><td></td><td></td><td></td><td> 110</td><td></td><td></td>

<td>val</td><td>Phe</td><td>Pro</td><td>Leu</td><td>Ala</td><td>Pro</td><td>cys</td><td>Cheese</td><td>Arg</td><td>Cheese</td><td>Thr</td><td>Cheese</td><td>Glu</td><td>Cheese</td><td>Thr</td><td>Ala</td>

<td></td><td></td><td> 115</td><td></td><td></td><td></td><td></td><td> 120</td><td></td><td></td><td></td><td></td><td> 125</td><td></td><td></td><td></td>

<td>Ala</td><td>Leu</td><td>Gly</td><td>Cys</td><td>Leu</td><td>Val</td><td>Lys</td><td>Asp</td><td>Tyr</td><td>Phe</td><td>Pro</td><td>Glu</td><td>Pro</td><td>Val</td><td>Thr</td><td>Val</td>

<td></td><td> 130</td><td></td><td></td><td></td><td></td><td> 135</td><td></td><td></td><td></td><td></td><td> 140</td><td></td><td></td><td></td><td></td>

<td>Cheese</td><td>Trp</td><td>Asn</td><td>Cheese</td><td>Gly</td><td>Ala</td><td>Leu</td><td>Thr</td><td>Cheese</td><td></td><td></td><td></td><td></td><td></td><td></td><td></td>

145 150 &lt;210&gt; 9 &lt;211&gt; 167 &lt;212&gt; PRT &lt;213&gt; Homo sapiens &lt;220&gt;

&lt;221&gt; MODJES &lt;222&gt; (103) &lt;223&gt; Any amino acid &lt;400&gt; 9

<td>Gly 1</td><td>Val</td><td>Val</td><td>Gin</td><td>Pro 5</td><td>Gly</td><td>Arg</td><td>Cheese</td><td>Leu</td><td>Arg 10</td><td>Leu</td><td>Cheese</td><td>Cys</td><td>Ala</td><td>Ala 15</td><td>Cheese</td>

<td>Gly</td><td>Phe</td><td>Thr</td><td>Phe twenty</td><td>Cheese</td><td>Cheese</td><td>Tyr</td><td>Gly</td><td>Underworld 25</td><td>His</td><td>Trp</td><td>Val</td><td>Arg</td><td>Gin thirty</td><td>Ala</td><td>Pro</td>

<td>Gly</td><td>Lys</td><td>Gly 35</td><td>Leu</td><td>Glu</td><td>Trp</td><td>Val</td><td>Ala 40</td><td>Val</td><td>How much</td><td>Trp</td><td>Tyr</td><td>Asp 45</td><td>Gly</td><td>Cheese</td><td>Asn</td>

<td>Lys</td><td>Tyr 50</td><td>Tyr</td><td>Ala</td><td>Asp</td><td>Cheese</td><td>Val 55</td><td>Lys</td><td>Gly</td><td>Arg</td><td>Phe</td><td>Thr 60</td><td>How much</td><td>Cheese</td><td>Arg</td><td>Asp</td>

Asn Ser Lys Asn Thr Leu Tyr Leu Gin Met Asn Ser Leu Arg Ala Glu

PL 214 003 Β1

<td> 65</td><td></td><td></td><td></td><td></td><td> 7 0</td><td></td><td></td><td></td><td></td><td> 75</td><td></td><td></td><td></td><td></td><td> 90</td>

<td>Asp</td><td>Thr</td><td>Ala</td><td>val</td><td>Tyr</td><td>Tyr</td><td>Cys</td><td>^ 11 a.</td><td>Arg</td><td>Asp</td><td>Pro</td><td>Arg</td><td>Gly</td><td>Ala</td><td>Thr</td><td>Leu</td>

<td></td><td></td><td></td><td></td><td> 85</td><td></td><td></td><td></td><td></td><td> 90</td><td></td><td></td><td></td><td></td><td> 95</td><td></td>

<td>Tyr</td><td>Tyr</td><td>Tyr</td><td>Tyr</td><td>Tyr</td><td>Arg</td><td>Xaa</td><td>Asp</td><td>Val</td><td>Trp</td><td>Gly</td><td>Gin</td><td>Gly</td><td>Thr</td><td>Thr</td><td>Val</td>

<td></td><td></td><td></td><td> 100</td><td></td><td></td><td></td><td></td><td> 105</td><td></td><td></td><td></td><td></td><td> 110</td><td></td><td></td>

<td>Thr</td><td>Val</td><td>Cheese</td><td>Cheese</td><td>Ala</td><td>Cheese</td><td>Thr</td><td>Lys</td><td>Gly</td><td>Pro</td><td>Cheese</td><td>Val</td><td>Phe</td><td>Pro</td><td>Leu</td><td>Ala</td>

<td></td><td></td><td> 115</td><td></td><td></td><td></td><td></td><td> 120</td><td></td><td></td><td></td><td></td><td> 125</td><td></td><td></td><td></td>

<td>Pro</td><td>Cys</td><td>Cheese</td><td>Arg</td><td>Cheese</td><td>Thr</td><td>Cheese</td><td>Glu</td><td>Cheese</td><td>Thr</td><td>Ala</td><td>Ala</td><td>Leu</td><td>Gly</td><td>Cys</td><td>Leu</td>

<td></td><td> 130</td><td></td><td></td><td></td><td></td><td> 135</td><td></td><td></td><td></td><td></td><td> 140</td><td></td><td></td><td></td><td></td>

<td>Val</td><td>Lys</td><td>Asp</td><td>Tyr</td><td>Phe</td><td>Pro</td><td>Glu</td><td>Pro</td><td>Val</td><td>Thr</td><td>V a 1</td><td>Cheese</td><td>Trp</td><td>Asn</td><td>Cheese</td><td>dy</td>

<td> 145</td><td></td><td></td><td></td><td></td><td> 150</td><td></td><td></td><td></td><td></td><td> 155</td><td></td><td></td><td></td><td></td><td> 160</td>

<td>Ala</td><td>Leu</td><td>Thr</td><td>Cheese</td><td>Gly</td><td>Val</td><td>His</td><td></td><td></td><td></td><td></td><td></td><td></td><td></td><td></td><td></td>

165 &lt;210&gt; 10 &lt;211&gt; 151 &lt;212&gt; PRT &lt;213&gt; Homo sapiens &lt;400&gt; 10

<td>Gly 1</td><td>Val</td><td>Val</td><td>Gin</td><td>Pro 5</td><td colspan="2">Gly Arg</td><td>Cheese</td><td>Leu</td><td>Arg 10</td><td>Leu</td><td colspan="2">Cheese Cys</td><td>Ala</td><td>Ala 15</td><td>Cheese</td>

<td>strength</td><td>Phe</td><td>Thr</td><td>Phe</td><td>Cheese</td><td>Cheese</td><td>Tyr</td><td>Gly</td><td>Underworld</td><td>His</td><td>Trp</td><td>Val</td><td>Arg</td><td>Gin</td><td>Ala</td><td>Pro</td>

<td></td><td></td><td></td><td> 20</td><td></td><td></td><td></td><td></td><td> 25</td><td></td><td></td><td></td><td></td><td> 30</td><td></td><td></td>

<td>Gly</td><td>Lys</td><td>Gly</td><td>Leu</td><td>Glu</td><td>Trp</td><td>Val</td><td>Ala</td><td>Val</td><td>How much</td><td>Trp</td><td>Tyr</td><td>Asp</td><td>Gly</td><td>Cheese</td><td>His</td>

<td></td><td></td><td> 35</td><td></td><td></td><td></td><td></td><td> 40</td><td></td><td></td><td></td><td></td><td> 45</td><td></td><td></td><td></td>

<td>Lys</td><td>Tyr</td><td>Tyr</td><td>Ala</td><td>Asp</td><td>Cheese</td><td>Val</td><td>Lys</td><td>Gly</td><td>Arg</td><td>Phe</td><td>Thr</td><td>How much</td><td>Cheese</td><td>Arg</td><td>Asp</td>

<td></td><td> 50</td><td></td><td></td><td></td><td></td><td> 55</td><td></td><td></td><td></td><td></td><td> 60</td><td></td><td></td><td></td><td></td>

<td>Asn</td><td>Cheese</td><td>Lys</td><td>Asn</td><td>Thr</td><td>Leu</td><td>Tyr</td><td>Leu</td><td>Gin</td><td>Underworld</td><td>Asn</td><td>Cheese</td><td>Leu</td><td>Arg</td><td>Al a</td><td>Glu</td>

<td> 65</td><td></td><td></td><td></td><td></td><td> 70</td><td></td><td></td><td></td><td></td><td> 75</td><td></td><td></td><td></td><td></td><td> 80</td>

<td>Asp</td><td>Thr</td><td>Ala</td><td>Val</td><td>Tyr</td><td>Tyr</td><td>Cys</td><td>Ala</td><td>Arg</td><td>Gly</td><td>Ala</td><td>Val</td><td>Val</td><td>Val</td><td>Pro</td><td>Ala</td>

<td></td><td></td><td></td><td></td><td> 85</td><td></td><td></td><td></td><td></td><td> 90</td><td></td><td></td><td></td><td></td><td> 95</td><td></td>

<td>Al a</td><td>Underworld</td><td>Asp</td><td>Val</td><td>Trp</td><td>Gly</td><td>Gin</td><td>Giy</td><td>Thr</td><td>Thr</td><td>Val</td><td>Thr</td><td>Val</td><td>Cheese</td><td>Cheese</td><td>Ala</td>

<td></td><td></td><td></td><td> 100</td><td></td><td></td><td></td><td></td><td> 105</td><td></td><td></td><td></td><td></td><td> 110</td><td></td><td></td>

<td>Cheese</td><td>Thr</td><td>Lys</td><td>Gly</td><td>Pro</td><td>Cheese</td><td>Val</td><td>Phe</td><td>Pro</td><td>Leu</td><td>Ala</td><td>Pro</td><td>Cys</td><td>Cheese</td><td>Arg</td><td>Cheese</td>

<td></td><td></td><td> 115</td><td></td><td></td><td></td><td></td><td> 120</td><td></td><td></td><td></td><td></td><td> 125</td><td></td><td></td><td></td>

<td>Thr</td><td>Cheese</td><td>Glu</td><td>Cheese</td><td>Thr</td><td>Ala</td><td>Ala</td><td>Leu</td><td>Gly</td><td>Cys</td><td>Leu</td><td>Val</td><td>Lys</td><td>Asp</td><td>Tyr</td><td>Phe</td>

130 135 140

Pro Glu Pro Val Thr Val Ser

145 150

PL 214 003 Β1

<td colspan="2">&lt;210&gt; 11 &lt;211&gt; 146 &lt;212&gt; PRT &lt;213&gt; Homo</td><td colspan="13">sapiens</td>

<td colspan="2"> &lt;220&gt;</td><td></td><td></td><td></td><td></td><td></td><td></td><td></td><td></td><td></td><td></td><td></td><td></td><td></td>

<td colspan="3">&lt;221&gt; MOD RES</td><td></td><td></td><td></td><td></td><td></td><td></td><td></td><td></td><td></td><td></td><td></td><td></td>

<td colspan="2"> &lt;222&gt; (22)</td><td></td><td></td><td></td><td></td><td></td><td></td><td></td><td></td><td></td><td></td><td></td><td></td><td></td>

<td colspan="2">&lt;223&gt; Any,</td><td colspan="3">new amino acid</td><td></td><td></td><td></td><td></td><td></td><td></td><td></td><td></td><td></td><td></td>

<td colspan="2"> &lt;400&gt; 11</td><td></td><td></td><td></td><td></td><td></td><td></td><td></td><td></td><td></td><td></td><td></td><td></td><td></td>

<td>Val</td><td>Val Gin</td><td>Pro</td><td>Gly</td><td>Arg</td><td>Cheese</td><td>Leu</td><td>Arg</td><td>Leu</td><td>Cheese</td><td>Cys</td><td>Ala</td><td>Ala</td><td>Cheese</td><td>Gly</td>

<td> 1</td><td></td><td></td><td> 5</td><td></td><td></td><td></td><td></td><td> 10</td><td></td><td></td><td></td><td></td><td> 15</td><td></td>

<td>Phe</td><td>Thr Phe</td><td>Cheese</td><td>Cheese</td><td>Xaa</td><td>Gly</td><td>Underworld</td><td>His</td><td>Trp</td><td>Val</td><td>Arg</td><td>Gin</td><td>Ala</td><td>Pro</td><td>Gly</td>

<td></td><td></td><td> 20</td><td></td><td></td><td></td><td></td><td> 25</td><td></td><td></td><td></td><td></td><td> 30</td><td></td><td></td>

<td>Lys</td><td>Gly Leu</td><td>Glu</td><td>Trp</td><td>Val</td><td>Ala</td><td>Val</td><td>How much</td><td>Trp</td><td>Cheese</td><td>Asp</td><td>Gly</td><td>Cheese</td><td>His</td><td>Bald</td>

<td></td><td> 35</td><td></td><td></td><td></td><td></td><td> 40</td><td></td><td></td><td></td><td></td><td> 45</td><td></td><td></td><td></td>

<td>Tyr</td><td>Tire Ala</td><td>Asp</td><td>Cheese</td><td>Val</td><td>Lys</td><td>Gly</td><td>Arg</td><td>Phe</td><td>Thr</td><td>how much</td><td>Cheese</td><td>Arg</td><td>Asp</td><td>Asn</td>

<td></td><td> 50</td><td></td><td></td><td></td><td> 55</td><td></td><td></td><td></td><td></td><td> 60</td><td></td><td></td><td></td><td></td>

<td>Cheese</td><td>Lys Asn</td><td>Thr</td><td>Leu</td><td>Tyr</td><td>Leu</td><td>Gin</td><td>Underworld</td><td>Asn</td><td>Cheese</td><td>Leu</td><td>Arg</td><td>Ala</td><td>Glu</td><td>Asp</td>

<td> 65</td><td></td><td></td><td></td><td> 70</td><td></td><td></td><td></td><td></td><td> 7 5</td><td></td><td></td><td></td><td></td><td> 80</td>

<td>Thr</td><td>Ala Val</td><td>Tyr</td><td>Tyr</td><td>Cys</td><td>Ala</td><td>Arg</td><td>Gly</td><td>Thr</td><td>Underworld</td><td>How much</td><td>Val</td><td>Val</td><td>Games</td><td>Thr</td>

<td></td><td></td><td></td><td> 85</td><td></td><td></td><td></td><td></td><td> 90</td><td></td><td></td><td></td><td></td><td> 95</td><td></td>

<td>Leu</td><td>Asp Tyr</td><td>Trp</td><td>Gly</td><td>Gin</td><td>Gly</td><td>Thr</td><td>Leu</td><td>Val</td><td>Thr</td><td>Val</td><td>Cheese</td><td>Cheese</td><td>Ala</td><td>Cheese</td>

<td></td><td></td><td> 100</td><td></td><td></td><td></td><td></td><td> 105</td><td></td><td></td><td></td><td></td><td> 110</td><td></td><td></td>

<td>Thr</td><td>Lys Gly</td><td>Pro</td><td>Cheese</td><td>Val</td><td>Phe</td><td>Pro</td><td>Leu</td><td>Ala</td><td>Pro</td><td>Cys</td><td>Cheese</td><td>Arg</td><td>Cheese</td><td>Thr</td>

<td></td><td> 115</td><td></td><td></td><td></td><td></td><td> 120</td><td></td><td></td><td></td><td></td><td> 125</td><td></td><td></td><td></td>

<td>Cheese</td><td>Glu Ser</td><td>Thr</td><td>Ala</td><td>Ala</td><td>Leu</td><td>Gly</td><td>Cys</td><td>Leu</td><td>Val</td><td>Lys</td><td>Asp</td><td>Tyr</td><td>Phe</td><td>Pro</td>

<td></td><td> 130</td><td></td><td></td><td></td><td> 135</td><td></td><td></td><td></td><td></td><td> 140</td><td></td><td></td><td></td><td></td>

<td>Glu</td><td>Pro</td><td></td><td></td><td></td><td></td><td></td><td></td><td></td><td></td><td></td><td></td><td></td><td></td><td></td>

<td> 145</td><td></td><td></td><td></td><td></td><td></td><td></td><td></td><td></td><td></td><td></td><td></td><td></td><td></td><td></td>

<td colspan="2"> &lt;210&gt; 12</td><td></td><td></td><td></td><td></td><td></td><td></td><td></td><td></td><td></td><td></td><td></td><td></td><td></td>

<td colspan="2"> &lt;211&gt; 174</td><td></td><td></td><td></td><td></td><td></td><td></td><td></td><td></td><td></td><td></td><td></td><td></td><td></td>

<td colspan="2">with 12 PRTs</td><td></td><td></td><td></td><td></td><td></td><td></td><td></td><td></td><td></td><td></td><td></td><td></td><td></td>

<td colspan="2">&lt;213&gt; Homo;</td><td colspan="2">sapiens</td><td></td><td></td><td></td><td></td><td></td><td></td><td></td><td></td><td></td><td></td><td></td>

<td colspan="2"> &lt;400&gt; 12</td><td></td><td></td><td></td><td></td><td></td><td></td><td></td><td></td><td></td><td></td><td></td><td></td><td></td>

<td>Cheese</td><td>Gly Gly</td><td>Gly</td><td>Val</td><td>vai</td><td>Gin</td><td>Pro</td><td>Gly</td><td>Arg</td><td>Cheese</td><td>Leu</td><td>Arg</td><td>Leu</td><td>Cheese</td><td>Cys</td>

<td> 1</td><td></td><td></td><td> 5</td><td></td><td></td><td></td><td></td><td> 10</td><td></td><td></td><td></td><td></td><td> 15</td><td></td>

<td>Ala</td><td>Ala Ser</td><td>Gly</td><td>Phe</td><td>Thr</td><td>Phe</td><td>Cheese</td><td>Cheese</td><td>Tyr</td><td>Gly</td><td>Val</td><td>His</td><td>Trp</td><td>Val</td><td>Arg</td>

<td></td><td></td><td> 20</td><td></td><td></td><td></td><td></td><td> 25</td><td></td><td></td><td></td><td></td><td> 30</td><td></td><td></td>

<td>Gin</td><td>Ala Pro</td><td>Gly</td><td>Lys</td><td>Gly</td><td>Leu</td><td>Glu</td><td>Trp</td><td>Val</td><td>Ala</td><td>Val</td><td>How much</td><td>Trp</td><td>Tyr</td><td>Asp</td>

<td></td><td> 35</td><td></td><td></td><td></td><td></td><td> 40</td><td></td><td></td><td></td><td></td><td> 45</td><td></td><td></td><td></td>

<td>Gly</td><td>Asn cheese</td><td>Lys</td><td>Tyr</td><td>Tyr</td><td>Ala</td><td>Asp</td><td>Cheese</td><td>Val</td><td>Lys</td><td>Gly</td><td>Arg</td><td>Phe</td><td>Thr</td><td>He</td>

PL 214 003 Β1

<td colspan="5"> 50</td><td colspan="5"> 55</td><td colspan="6"> 60</td>

<td>Cheese</td><td>Arg</td><td>Asp</td><td>Asn</td><td>Cheese</td><td>Lys</td><td>Cheese</td><td>Thr</td><td>Leu</td><td>Tyr</td><td>Leu</td><td>Gin</td><td>Underworld</td><td>Asn</td><td>Cheese</td><td>Leu</td>

<td> 65</td><td></td><td></td><td></td><td></td><td> 70</td><td></td><td></td><td></td><td></td><td> 75</td><td></td><td></td><td></td><td></td><td> 00</td>

<td>Arg</td><td>Ala</td><td>Glu</td><td>Asp</td><td>Thr</td><td>Ala</td><td>Val</td><td>Tyr</td><td>Tyr</td><td>Cys</td><td>Ala</td><td>Arg</td><td>Asp</td><td>Cheese</td><td>Tyr</td><td>Tyr</td>

<td></td><td></td><td></td><td></td><td> 85</td><td></td><td></td><td></td><td></td><td> 90</td><td></td><td></td><td></td><td></td><td> 95</td><td></td>

<td>Asp</td><td>Phe</td><td>Trp</td><td>Cheese</td><td>Gly</td><td>Arg</td><td>Gly</td><td>Gly</td><td>Underworld</td><td>Asp</td><td>Val</td><td>Trp</td><td>Gly</td><td>Gin</td><td>Gly</td><td>Thr</td>

<td></td><td></td><td></td><td> 100</td><td></td><td></td><td></td><td></td><td> 105</td><td></td><td></td><td></td><td></td><td> 110</td><td></td><td></td>

<td>Thr</td><td>Val</td><td>Thr</td><td>Val</td><td>Cheese</td><td>Cheese</td><td>Ala</td><td>Cheese</td><td>Thr</td><td>Lys</td><td>Gly</td><td>Pro</td><td>Cheese</td><td>val</td><td>Phe</td><td>Pro</td>

<td></td><td></td><td> 115</td><td></td><td></td><td></td><td></td><td> 120</td><td></td><td></td><td></td><td></td><td> 12 5</td><td></td><td></td><td></td>

<td>Leu</td><td>Ala</td><td>Pro</td><td>Cys</td><td>Cheese</td><td>Arg</td><td>Cheese</td><td>Thr</td><td>Cheese</td><td>Glu</td><td>Cheese</td><td>Thr</td><td>Ala</td><td>Ala</td><td>Leu</td><td>c-iy</td>

<td></td><td> 130</td><td></td><td></td><td></td><td></td><td> 135</td><td></td><td></td><td></td><td></td><td> 140</td><td></td><td></td><td></td><td></td>

<td>Cys</td><td>Leu</td><td>Val</td><td>Lys</td><td>Asp</td><td>Tyr</td><td>Phe</td><td>Pro</td><td>Glu</td><td>Pro</td><td>Val</td><td>Thr</td><td>Val</td><td>Cheese</td><td>Trp</td><td>Asn</td>

<td> 145</td><td></td><td></td><td></td><td></td><td>1 by 0</td><td></td><td></td><td></td><td></td><td> 155</td><td></td><td></td><td></td><td></td><td> 160</td>

<td>Cheese</td><td>G1 y</td><td>Ala</td><td>JLe u</td><td>Thr</td><td>Cheese</td><td>Gly</td><td>Val</td><td>His</td><td>Thr</td><td>Phe</td><td>Pro</td><td>Ala</td><td>Val</td><td></td><td></td>

170 &lt;210&gt; 13 &lt;211&gt; 163 &lt;212&gt; PRT &lt;213&gt; Homo sapiens

<td colspan="16"> &lt;400&gt; 13</td>

<td>Val 1</td><td>Gin</td><td>Pro</td><td colspan="2">Gly Arg 5</td><td colspan="2">Leu cheese</td><td>Arg</td><td>Leu</td><td>Cheese 10</td><td>Cys</td><td>Ala</td><td>Ala</td><td>Cheese</td><td>Gly 15</td><td>Phe</td>

<td>Thr</td><td>Phe</td><td>Cheese</td><td>Asn</td><td>Tyr</td><td>Ala</td><td>Underworld</td><td>His</td><td>Trp</td><td>Val</td><td>Arg</td><td>G1 n</td><td>Ala</td><td>Pro</td><td>Gly</td><td>Lys</td>

<td></td><td></td><td></td><td> 2 0</td><td></td><td></td><td></td><td></td><td> 25</td><td></td><td></td><td></td><td></td><td> 30</td><td></td><td></td>

<td>Gly</td><td>Leu</td><td>Glu</td><td>Trp</td><td>Val</td><td>Val</td><td>Val</td><td>Background</td><td>_Trp</td><td>His</td><td>Asp</td><td>Gly</td><td>Asn</td><td>Asn</td><td>Lys</td><td>Tyr</td>

<td></td><td></td><td> 35</td><td></td><td></td><td></td><td></td><td> 40</td><td></td><td></td><td></td><td></td><td> 45</td><td></td><td></td><td></td>

<td>Tyr</td><td>A .. a</td><td>Glu</td><td>Cheese</td><td>Val</td><td>Lys</td><td>Gly</td><td>Arg</td><td>Phe</td><td>Thr</td><td>How much</td><td>Cheese</td><td>Arg</td><td>Asp</td><td>Asn</td><td>Cheese</td>

<td></td><td> 50</td><td></td><td></td><td></td><td></td><td> 55</td><td></td><td></td><td></td><td></td><td> 60</td><td></td><td></td><td></td><td></td>

<td>Lys</td><td>Asn</td><td>Thr</td><td>Leu</td><td>Tyr</td><td>Leu</td><td>Gin</td><td>Underworld</td><td>Asn</td><td>Cheese</td><td>Leu</td><td>Arg</td><td>Ala</td><td>Glu</td><td>Asp</td><td>Thr</td>

<td> 65</td><td></td><td></td><td></td><td></td><td> 70</td><td></td><td></td><td></td><td></td><td> 75</td><td></td><td></td><td></td><td></td><td> 80</td>

<td>Ala</td><td>Val</td><td>Tyr</td><td>Tyr</td><td>Cys</td><td>Ala</td><td>Arg</td><td>Asp</td><td>Gin</td><td>Gly</td><td>Thr</td><td>Gly</td><td>Trp</td><td>Tyr</td><td>Gly</td><td>Gly</td>

<td></td><td></td><td></td><td></td><td> 85</td><td></td><td></td><td></td><td></td><td> 90</td><td></td><td></td><td></td><td></td><td> 95</td><td></td>

<td>Phe</td><td>Asp</td><td>Phe</td><td>Trp</td><td>Gly</td><td>Gin</td><td>Gly</td><td>Thr</td><td>Leu</td><td>Val</td><td>Thr</td><td>Val</td><td>Cheese</td><td>Cheese</td><td>Ala</td><td>Cheese</td>

<td></td><td></td><td></td><td> 100</td><td></td><td></td><td></td><td></td><td> 105</td><td></td><td></td><td></td><td></td><td> 110</td><td></td><td></td>

<td>Thr</td><td>Lys</td><td>Gly</td><td>Pro</td><td>Cheese</td><td>Val</td><td>Phe</td><td>Pro</td><td>Leu</td><td>Ala</td><td>Pro</td><td>Cys</td><td>Cheese</td><td>Arg</td><td>Cheese</td><td>Thr</td>

<td></td><td></td><td> 115</td><td></td><td></td><td></td><td></td><td> 120</td><td></td><td></td><td></td><td></td><td> 125</td><td></td><td></td><td></td>

<td>Cheese</td><td>Glu</td><td>Cheese</td><td>Thr</td><td>Ala</td><td>Ala</td><td>Leu</td><td>Gly</td><td>Cys</td><td>Leu</td><td>Val</td><td>Lys</td><td>Asp</td><td>Tyr</td><td>Phe</td><td>Pro</td>

<td></td><td> 130</td><td></td><td></td><td></td><td></td><td> 135</td><td></td><td></td><td></td><td></td><td> 140</td><td></td><td></td><td></td><td></td>

<td>Glu</td><td>Pro</td><td>Val</td><td>Thr</td><td>Val</td><td>Cheese</td><td>Trp</td><td>Asn</td><td>Cheese</td><td>Gly</td><td>Ala</td><td>Leu</td><td>Thr</td><td>Cheese</td><td>Gly</td><td>Val</td>

PL 214 003 Β1

145 150

His Thr Phe

155

160 &lt;210&gt; 14 &lt;211&gt; 235 &lt;212&gt; PRT &lt;213&gt; Homo sapiens &lt;400&gt; 14

<td>Underworld 1</td><td>Glu</td><td>Thr</td><td>Pro</td><td>Ala 5</td><td>Gin</td><td>Leu</td><td>Leu</td><td>Phe</td><td>Leu 10</td><td>Leu</td><td>Leu</td><td>Leu</td><td>Trp</td><td>Leu 15</td><td>Pro</td>

<td>Asp</td><td>Thr</td><td>Thr</td><td>Gly</td><td>Glu</td><td>How much</td><td>Val</td><td>Leu</td><td>Thr</td><td>Gin</td><td>Cheese</td><td>Pro</td><td>Gly</td><td>Thr</td><td>Leu</td><td>Cheese</td>

<td></td><td></td><td></td><td> 20</td><td></td><td></td><td></td><td></td><td> 25</td><td></td><td></td><td></td><td></td><td> 30</td><td></td><td></td>

<td>Leu</td><td>Cheese</td><td>Pro</td><td>Gly</td><td>Glu</td><td>Arg</td><td>. 1. cx</td><td>Thr</td><td>Leu</td><td>Cheese</td><td>cys</td><td>Arg</td><td>Ala</td><td>Cheese</td><td>Gin</td><td>Cheese</td>

<td></td><td></td><td> 35</td><td></td><td></td><td></td><td></td><td> 40</td><td></td><td></td><td></td><td></td><td> 45</td><td></td><td></td><td></td>

<td>How much</td><td>Cheese</td><td>Cheese</td><td>Cheese</td><td>Phe</td><td>Leu</td><td>Ala</td><td>Trp</td><td>Tyr</td><td>Gin</td><td>Gin</td><td>Arg</td><td>Pro</td><td>Gly</td><td>Gin</td><td>Ala</td>

<td></td><td> 50</td><td></td><td></td><td></td><td></td><td> 55</td><td></td><td></td><td></td><td></td><td> 60</td><td></td><td></td><td></td><td></td>

<td>Pro</td><td>Arg</td><td>Leu</td><td>Leu</td><td>How much</td><td>Tyr</td><td>Gly</td><td>Ala</td><td>Cheese</td><td>Cheese</td><td>Arg</td><td>Ala</td><td>Thr</td><td>Gly</td><td>How much</td><td>Pro</td>

<td> 65</td><td></td><td></td><td></td><td></td><td> 70</td><td></td><td></td><td></td><td></td><td> 7 5</td><td></td><td></td><td></td><td></td><td> 80</td>

<td>Asp</td><td>Arg</td><td>Phe</td><td>Cheese</td><td>Gly</td><td>Cheese</td><td>Gly</td><td>Cheese</td><td>Gly</td><td>Thr</td><td>Asp</td><td>Phe</td><td>Thr</td><td>Leu</td><td>Thr</td><td>How much</td>

<td></td><td></td><td></td><td></td><td> 85</td><td></td><td></td><td></td><td></td><td> 90</td><td></td><td></td><td></td><td></td><td> 95</td><td></td>

<td>Cheese</td><td>Arg</td><td>Leu</td><td>Glu</td><td>.Pro</td><td>Glu</td><td>Asp</td><td>Phe</td><td>Ala</td><td>Val</td><td>Tyr</td><td>Tyr</td><td>Cys</td><td>Gin</td><td>Gin</td><td>Tyr</td>

<td></td><td></td><td></td><td> 100</td><td></td><td></td><td></td><td></td><td> 105</td><td></td><td></td><td></td><td></td><td> 110</td><td></td><td></td>

<td>Gly</td><td>Thr</td><td>Cheese</td><td>Pro</td><td>Trp</td><td>Thr</td><td>Phe</td><td>Gly</td><td>Gin</td><td>Giy</td><td>Thr</td><td>Lys</td><td>Val</td><td>Glu</td><td>How much</td><td>Lys</td>

<td></td><td></td><td> 115</td><td></td><td></td><td></td><td></td><td> 120</td><td></td><td></td><td></td><td></td><td> 125</td><td></td><td></td><td></td>

<td>Arg</td><td>Thr</td><td>Val.</td><td>Ala</td><td>Ala</td><td>Pro</td><td>Cheese</td><td>Val</td><td>Phe</td><td>How much</td><td>Phe</td><td>Pro</td><td>Pro</td><td>Cheese</td><td>Asp</td><td>G1 u</td>

<td></td><td> 130</td><td></td><td></td><td></td><td></td><td> 135</td><td></td><td></td><td></td><td></td><td> 14 0</td><td></td><td></td><td></td><td></td>

<td>Gin</td><td>Leu</td><td>Lys</td><td>Cheese</td><td>Gly</td><td>Thr</td><td>, j. ci</td><td>Cheese</td><td>Val</td><td>Val</td><td>Cys</td><td>Leu</td><td>Leu</td><td>Asn</td><td>Asn</td><td>Phe</td>

<td> 145</td><td></td><td></td><td></td><td></td><td> 150</td><td></td><td></td><td></td><td></td><td> 155</td><td></td><td></td><td></td><td></td><td> 160</td>

<td>Tyr</td><td>Pro</td><td>Arg</td><td>Glu</td><td>Ala</td><td>Lys</td><td>Val</td><td>Gin</td><td>Trp</td><td>Lys</td><td>Val</td><td>Asp</td><td>Asn</td><td>Ala</td><td>Leu</td><td>Gin</td>

<td></td><td></td><td></td><td></td><td> 165</td><td></td><td></td><td></td><td></td><td> 170</td><td></td><td></td><td></td><td></td><td>2 * y</td><td></td>

<td>Cheese</td><td>Gly</td><td>Asn</td><td>Cheese</td><td>Gin</td><td>Glu</td><td>Cheese</td><td>val</td><td>Thr</td><td>Glu</td><td>Gin</td><td>Asp</td><td>Cheese</td><td>Lys</td><td>Asp</td><td>Cheese</td>

<td></td><td></td><td></td><td> 180</td><td></td><td></td><td></td><td></td><td> 185</td><td></td><td></td><td></td><td></td><td> 190</td><td></td><td></td>

<td>Thr</td><td>Tyr</td><td>Cheese</td><td>Leu</td><td>Cheese</td><td>Cheese</td><td>Thr</td><td>Leu</td><td>Thr</td><td>Leu</td><td>Cheese</td><td>Lys</td><td>Ala</td><td>Asp</td><td>Tyr</td><td>Glu</td>

<td></td><td></td><td> 195</td><td></td><td></td><td></td><td></td><td> 200</td><td></td><td></td><td></td><td></td><td> 205</td><td></td><td></td><td></td>

<td>Lys</td><td>His</td><td>Lys</td><td>Val</td><td>Tyr</td><td>Ala</td><td>Cys</td><td>Glu</td><td>Val</td><td>Thr</td><td>His</td><td>Gin</td><td>Gly</td><td>Leu</td><td>Cheese</td><td>Cheese</td>

<td></td><td> 210</td><td></td><td></td><td></td><td></td><td> 215</td><td></td><td></td><td></td><td></td><td> 220</td><td></td><td></td><td></td><td></td>

<td>Pro</td><td>Val</td><td>Thr</td><td>Lys</td><td>Cheese</td><td>Phe</td><td>Asn</td><td>Arg</td><td>Gly</td><td>Glu</td><td>Cys</td><td></td><td></td><td></td><td></td><td></td>

<td> 225</td><td></td><td></td><td></td><td></td><td> 230</td><td></td><td></td><td></td><td></td><td> 235</td><td></td><td></td><td></td><td></td><td></td>

&lt;210 15

PL 214 003 Β1 &lt;211&gt; 233 &lt;212&gt; PRT &lt;213&gt; Homo sapiens

<td colspan="6"> &lt;400&gt; 15</td><td rowspan="2">Leu</td><td rowspan="2">Leu</td><td rowspan="2">Phe</td><td rowspan="2">Leu 10</td><td colspan="3" rowspan="2">Leu Leu Leu</td><td rowspan="2">Trp</td><td rowspan="2">Leu 15</td><td rowspan="2">Pro</td>

<td>Underworld 1</td><td>Glu</td><td>Thr</td><td colspan="2">Pro Ala 5</td><td>Gin</td>

<td>Asp</td><td>Thr</td><td>Thr</td><td>Gly</td><td>Glu</td><td>how much</td><td>Val</td><td>Leu</td><td>Thr</td><td>Gin</td><td>Cheese</td><td>Pro</td><td>Gly</td><td>Thr</td><td>Leu</td><td>Cheese</td>

<td></td><td></td><td></td><td> 20</td><td></td><td></td><td></td><td></td><td> 25</td><td></td><td></td><td></td><td></td><td> 30</td><td></td><td></td>

<td>Leu</td><td>Cheese</td><td>Pro</td><td>Gly</td><td>Glu</td><td>Arg</td><td>Ala</td><td>Thr</td><td>Leu</td><td>Cheese</td><td>Cys</td><td>Arg</td><td>Thr</td><td>Cheese</td><td>Vai</td><td>Cheese</td>

<td></td><td></td><td> 35</td><td></td><td></td><td></td><td></td><td> 40</td><td></td><td></td><td></td><td></td><td> 45</td><td></td><td></td><td></td>

<td>Cheese</td><td>Cheese</td><td>Tyr</td><td>Leu</td><td>Ala</td><td>Trp</td><td>Tyr</td><td>Gin</td><td>Gin</td><td>Lys</td><td>Pro</td><td>Gly</td><td>Gin</td><td>Al a</td><td>Pro</td><td>Arg</td>

<td></td><td> 50</td><td></td><td></td><td></td><td></td><td> 55</td><td></td><td></td><td></td><td></td><td> 60</td><td></td><td></td><td></td><td></td>

<td>Leu</td><td>Leu</td><td>How much</td><td>Tyr</td><td>Gly</td><td>Ala</td><td>Cheese</td><td>Cheese</td><td>Arg</td><td>Ala</td><td>Thr</td><td>Gly</td><td>how much</td><td>Pro</td><td>Asp</td><td>Arg</td>

<td> 65</td><td></td><td></td><td></td><td></td><td> 70</td><td></td><td></td><td></td><td></td><td>η</td><td></td><td></td><td></td><td></td><td> 30</td>

<td>Phe</td><td>Cheese</td><td>Gly</td><td>Cheese</td><td>Gly</td><td>Cheese</td><td>Gly</td><td>Thr</td><td>Asp</td><td>Phe</td><td>Thr</td><td>Leu</td><td>Thr</td><td>How much</td><td>Cheese</td><td>Arg</td>

<td></td><td></td><td></td><td></td><td> 85</td><td></td><td></td><td></td><td></td><td> 90</td><td></td><td></td><td></td><td></td><td> 95</td><td></td>

<td>Leu</td><td>Glu</td><td>Pro</td><td>Glu</td><td>Asp</td><td>Phe</td><td>Ala</td><td>Val</td><td>Tyr</td><td>Tyr</td><td>Cys</td><td>Gin</td><td>Gin</td><td>Tyr</td><td>Gly</td><td>How much</td>

<td></td><td></td><td></td><td> 100</td><td></td><td></td><td></td><td></td><td> 105</td><td></td><td></td><td></td><td></td><td> 110</td><td></td><td></td>

<td>Cheese</td><td>Pro</td><td>Phe</td><td>Thr</td><td>Phe</td><td>Gly</td><td>Gly</td><td>Gly</td><td>Thr</td><td>Lys</td><td>Val</td><td>Glu</td><td>How much</td><td>Lys</td><td>Arg</td><td>Thr</td>

<td></td><td></td><td> 115</td><td></td><td></td><td></td><td></td><td> 120</td><td></td><td></td><td></td><td></td><td> 125</td><td></td><td></td><td></td>

<td>Val</td><td>Ala</td><td>Ala</td><td>Pro</td><td>Cheese</td><td>Val</td><td>Phe</td><td>How much</td><td>Phe</td><td>Pro</td><td>Pro</td><td>Cheese</td><td>Asp</td><td>Glu</td><td>Gin</td><td>Leu</td>

<td></td><td> 130</td><td></td><td></td><td></td><td></td><td> 135</td><td></td><td></td><td></td><td></td><td> 140</td><td></td><td></td><td></td><td></td>

<td>Lys</td><td>Cheese</td><td>Gly</td><td>Thr</td><td>Ala</td><td>Cheese</td><td>Val</td><td>Val</td><td>Cys</td><td>Leu</td><td>Leu</td><td>Asn</td><td>Asn</td><td>Phe</td><td>Tyr</td><td>Pro</td>

<td> 145</td><td></td><td></td><td></td><td></td><td> 150</td><td></td><td></td><td></td><td></td><td> 155</td><td></td><td></td><td></td><td></td><td> 160</td>

<td>Arg</td><td>Glu</td><td>Ala</td><td>Lys</td><td>Val</td><td>Gin</td><td>Trp</td><td>Lys</td><td>val</td><td>Asp</td><td>Asn</td><td>Ala</td><td>Leu</td><td>Gin</td><td>Cheese</td><td>Gly</td>

<td></td><td></td><td></td><td></td><td> 165</td><td></td><td></td><td></td><td></td><td> 170</td><td></td><td></td><td></td><td></td><td> 175</td><td></td>

<td>Asn</td><td>Cheese</td><td>Gin</td><td>GXu</td><td>Cheese</td><td>vai</td><td>Thr</td><td>Glu</td><td>Gin</td><td>Asp</td><td>Cheese</td><td>Lys</td><td>Asp</td><td>Cheese</td><td>Thr</td><td>Tyr</td>

<td></td><td></td><td></td><td> 180</td><td></td><td></td><td></td><td></td><td> 185</td><td></td><td></td><td></td><td></td><td> 190</td><td></td><td></td>

<td>Cheese</td><td>Leu</td><td>Cheese</td><td>Cheese</td><td>Thr</td><td>Leu</td><td>Thr</td><td>Leu</td><td>Cheese</td><td>Lys</td><td>Ala</td><td>Asp</td><td>Tyr</td><td>Glu</td><td>Lys</td><td>His</td>

<td></td><td></td><td> 195</td><td></td><td></td><td></td><td></td><td> 200</td><td></td><td></td><td></td><td></td><td> 205</td><td></td><td></td><td></td>

<td>Lys</td><td>Val</td><td>Tyr</td><td>7X1 S.</td><td>Cys</td><td>Glu</td><td>Val</td><td>Thr</td><td>His</td><td>Gin</td><td>Gly</td><td>Leu</td><td>Cheese</td><td>Cheese</td><td>Pro</td><td>Val</td>

<td></td><td> 210</td><td></td><td></td><td></td><td></td><td> 215</td><td></td><td></td><td></td><td></td><td> 220</td><td></td><td></td><td></td><td></td>

<td>Thr</td><td>Lys</td><td>Cheese</td><td>Phe</td><td>Asn</td><td>Arg</td><td>Gly</td><td>Glu</td><td>Cys</td><td></td><td></td><td></td><td></td><td></td><td></td><td></td>

<td> 225</td><td></td><td></td><td></td><td></td><td> 230</td><td></td><td></td><td></td><td></td><td></td><td></td><td></td><td></td><td></td><td></td>

&lt;210&gt; 16 &lt;211&gt; 139 &lt;212&gt; PRT &lt;213&gt; Homo sapiens &lt;400&gt; 16

Gly Thr Leu Ser Leu Ser Pro Gly Glu Arg Ala Thr Leu Ser Cys Arg

PL 214 003 Β1

<td> 1</td><td></td><td></td><td></td><td> 5</td><td></td><td></td><td></td><td></td><td> 10</td><td></td><td></td><td></td><td></td><td> 15</td><td></td>

<td>AJ. those</td><td>Cheese</td><td>Gin</td><td>Cheese</td><td>vai</td><td>Cheese</td><td>Cheese</td><td>Tyr</td><td>Leu</td><td>Ala</td><td>Trp</td><td>Tyr</td><td>Gin</td><td>Gin</td><td>Lys</td><td>Pro</td>

<td></td><td></td><td></td><td> 20</td><td></td><td></td><td></td><td></td><td> 25</td><td></td><td></td><td></td><td></td><td> 30</td><td></td><td></td>

<td>Gly</td><td>Gin</td><td>Ala</td><td>Pro</td><td>Arg</td><td>Leu</td><td>Leu</td><td>How much</td><td>Tyr</td><td>Gly</td><td>Ala</td><td>Cheese</td><td>Cheese</td><td>Arg</td><td>Ala</td><td>Thr</td>

<td></td><td></td><td> 35</td><td></td><td></td><td></td><td></td><td> 40</td><td></td><td></td><td></td><td></td><td> 45</td><td></td><td></td><td></td>

<td>dy</td><td>How much</td><td>Pro</td><td>Asp</td><td>Arg</td><td>Phe</td><td>Cheese</td><td>Gly</td><td>Cheese</td><td>Gly</td><td>Cheese</td><td>Gly</td><td>Thr</td><td>Asp</td><td>Phe</td><td>Thr</td>

<td></td><td> 50</td><td></td><td></td><td></td><td></td><td> 55</td><td></td><td></td><td></td><td></td><td> 60</td><td></td><td></td><td></td><td></td>

<td>Leu</td><td>Thr</td><td>How much</td><td>Cheese</td><td>Arg</td><td>Leu</td><td>Glu</td><td>Pro</td><td>Glu</td><td>Asp</td><td>Phe</td><td>Ala</td><td>val</td><td>Tyr</td><td>Tyr</td><td>Cys</td>

<td> 65</td><td></td><td></td><td></td><td></td><td> 70</td><td></td><td></td><td></td><td></td><td> 75</td><td></td><td></td><td></td><td></td><td> 80</td>

<td>Gin</td><td>Gin</td><td>Tyr</td><td>Gly</td><td>Arg</td><td>Cheese</td><td>Pro</td><td>Phe</td><td>Thr</td><td>Phe</td><td>Gly</td><td>Pro</td><td>Gly</td><td>Thr</td><td>Lys</td><td>vai</td>

<td></td><td></td><td></td><td></td><td> 85</td><td></td><td></td><td></td><td></td><td> 90</td><td></td><td></td><td></td><td></td><td> 95</td><td></td>

<td>Asp</td><td>He</td><td>Lys</td><td>Arg</td><td>Thr</td><td>Val</td><td>Ala</td><td>Ala</td><td>Pro</td><td>Cheese</td><td>vai</td><td>Phe</td><td>How much</td><td>Phe</td><td>Pro</td><td>Pro</td>

<td></td><td></td><td></td><td> 100</td><td></td><td></td><td></td><td></td><td> 105</td><td></td><td></td><td></td><td></td><td> 110</td><td></td><td></td>

<td>Cheese</td><td>Asp</td><td>Glu</td><td>Gin</td><td>Leu</td><td>Lys</td><td>Cheese</td><td>Gly</td><td>Thr</td><td>Ala</td><td>Cheese</td><td>Val</td><td>Val</td><td>Cys</td><td>Leu</td><td>Leu</td>

<td></td><td></td><td> 115</td><td></td><td></td><td></td><td></td><td> 120</td><td></td><td></td><td></td><td></td><td> 125</td><td></td><td></td><td></td>

<td>Asn</td><td>Asn</td><td>Phe</td><td>Tyr</td><td>Pro</td><td>Arg</td><td>Glu</td><td>Ala</td><td>Lys</td><td>Val</td><td>Gin</td><td></td><td></td><td></td><td></td><td></td>

130 135 &lt;210&gt; 17 &lt;211&gt; 234 &lt;212&gt; PRT &lt;213&gt; Homo sapiens &lt;400&gt; 17

<td>Underworld 1</td><td>Glu</td><td>Thr</td><td>Pro</td><td>Ala 5</td><td>Gin</td><td colspan="2">Leu Leu</td><td>Phe</td><td>Leu 10</td><td>Leu</td><td>Leu</td><td>Leu</td><td>Trp</td><td>Leu 15</td><td>Pro</td>

<td>Asp</td><td>Thr</td><td>Thr</td><td>Gly</td><td>Glu</td><td>How much</td><td>Val</td><td>Leu</td><td>Thr</td><td>Gin</td><td>Cheese</td><td>Pro</td><td>Gly</td><td>Thr</td><td>Leu</td><td>Cheese</td>

<td></td><td></td><td></td><td> 20</td><td></td><td></td><td></td><td></td><td> 25</td><td></td><td></td><td></td><td></td><td> 30</td><td></td><td></td>

<td>Leu</td><td>Cheese</td><td>Pro</td><td>Gly</td><td>Glu</td><td>Arg</td><td>Ala</td><td>Thr</td><td>Leu</td><td>Cheese</td><td>Cys</td><td>Arg</td><td>Ala</td><td>Cheese</td><td>Gin</td><td>Cheese</td>

<td></td><td></td><td> 35</td><td></td><td></td><td></td><td></td><td> 40</td><td></td><td></td><td></td><td></td><td> 45</td><td></td><td></td><td></td>

<td>Val</td><td>Cheese</td><td>Cheese</td><td>Tyr</td><td>Leu</td><td>Ala</td><td>Trp</td><td>Tyr</td><td>Gin</td><td>Gin</td><td>Lys</td><td>Pro</td><td>Gly</td><td>Gin</td><td>Ala</td><td>Pro</td>

<td></td><td> 50</td><td></td><td></td><td></td><td></td><td> 55</td><td></td><td></td><td></td><td></td><td> 60</td><td></td><td></td><td></td><td></td>

<td>Arg</td><td>Pro</td><td>Leu</td><td>How much</td><td>Tyr</td><td>dy</td><td>Val</td><td>Cheese</td><td>Cheese</td><td>Arg</td><td>Ala</td><td>Thr</td><td>Gly</td><td>How much</td><td>Pro</td><td>Asp</td>

<td> 65</td><td></td><td></td><td></td><td></td><td> 70</td><td></td><td></td><td></td><td></td><td> 75</td><td></td><td></td><td></td><td></td><td> 80</td>

<td>Arg</td><td>Phe</td><td>Cheese</td><td>Gly</td><td>Cheese</td><td>Gly</td><td>Cheese</td><td>Gly</td><td>Thr</td><td>Asp</td><td>Phe</td><td>Thr</td><td>Leu</td><td>Thr</td><td>How much</td><td>Cheese</td>

<td></td><td></td><td></td><td></td><td> 85</td><td></td><td></td><td></td><td></td><td> 90</td><td></td><td></td><td></td><td></td><td> 95</td><td></td>

<td>Arg</td><td>Leu</td><td>Glu</td><td>Pro</td><td>Glu</td><td>Asp</td><td>Phe</td><td>Ala</td><td>Val</td><td>Tyr</td><td>Tyr</td><td>Cys</td><td>Gin</td><td>Gin</td><td>Tyr</td><td>Gly</td>

<td></td><td></td><td></td><td> 100</td><td></td><td></td><td></td><td></td><td> 105</td><td></td><td></td><td></td><td></td><td> 110</td><td></td><td></td>

<td>How much</td><td>Cheese</td><td>Pro</td><td>Phe</td><td>Thr</td><td>Phe</td><td>dy</td><td>Pro</td><td>Gly</td><td>Thr</td><td>Lys</td><td>Val</td><td>Asp</td><td>How much</td><td>Lys</td><td>Arg</td>

<td></td><td></td><td> 115</td><td></td><td></td><td></td><td></td><td> 120</td><td></td><td></td><td></td><td></td><td> 125</td><td></td><td></td><td></td>

<td>Thr</td><td>Val</td><td>Ala</td><td>Ala</td><td>Pro</td><td>Cheese</td><td>Val</td><td>Phe</td><td>How much</td><td>Phe</td><td>Pro</td><td>Pro</td><td>Cheese</td><td>Asp</td><td>Glu</td><td>Gin</td>

PL 214 003 Β1

<td></td><td> 130</td><td></td><td></td><td></td><td></td><td> 135</td><td></td><td></td><td></td><td></td><td> 140</td><td></td><td></td><td></td><td></td>

<td>Leu</td><td>Lys</td><td>Cheese</td><td>Gly</td><td>Thr</td><td>Ala</td><td>Cheese</td><td>Val</td><td>Val</td><td>Cys</td><td>Leu</td><td>Leu</td><td>Asn</td><td>Asn</td><td>Phe</td><td>Tyr</td>

<td> 145</td><td></td><td></td><td></td><td></td><td> 150</td><td></td><td></td><td></td><td></td><td> 155</td><td></td><td></td><td></td><td></td><td> 160</td>

<td>Pro</td><td>Arg</td><td>Glu</td><td>Ala</td><td>Lys</td><td>Val</td><td>Gin</td><td>Trp</td><td>Lys</td><td>Vai</td><td>Asp</td><td>Asn</td><td>Ala</td><td>Leu</td><td>Gin</td><td>Cheese</td>

<td></td><td></td><td></td><td></td><td> 165</td><td></td><td></td><td></td><td></td><td> 170</td><td></td><td></td><td></td><td></td><td> 175</td><td></td>

<td>Gly</td><td>Asn</td><td>Cheese</td><td>Gin</td><td>Glu</td><td>Cheese</td><td>Val</td><td>Thr</td><td>Glu</td><td>Gin</td><td>Asp</td><td>Cheese</td><td>Lys</td><td>Asp</td><td>Cheese</td><td>Thr</td>

<td></td><td></td><td></td><td> 180</td><td></td><td></td><td></td><td></td><td> 185</td><td></td><td></td><td></td><td></td><td> 190</td><td></td><td></td>

<td>Tyr</td><td>Cheese</td><td>Leu</td><td>Cheese</td><td>Cheese</td><td>Thr</td><td>Leu</td><td>Thr</td><td>Leu</td><td>Cheese</td><td>Lys</td><td>Ala</td><td>Asp</td><td>Tyr</td><td>Glu</td><td>Lys</td>

<td></td><td></td><td> 195</td><td></td><td></td><td></td><td></td><td> 200</td><td></td><td></td><td></td><td></td><td> 205</td><td></td><td></td><td></td>

<td>His</td><td>Lys</td><td>Val</td><td>Tyr</td><td>Ala</td><td>cys</td><td>Glu</td><td>Val</td><td>Thr</td><td>His</td><td>Gin</td><td>Gly</td><td>Leu</td><td>Cheese</td><td>Cheese</td><td>Pro</td>

<td></td><td> 210</td><td></td><td></td><td></td><td></td><td> 215</td><td></td><td></td><td></td><td></td><td> 220</td><td></td><td></td><td></td><td></td>

<td>Val</td><td>Thr</td><td>Lys</td><td>Cheese</td><td>Phe</td><td>Asn</td><td>Arg</td><td>Gly</td><td>Glu</td><td>cys</td><td></td><td></td><td></td><td></td><td></td><td></td>

225 230 &lt;210&gt; 18 &lt;211&gt; 152 &lt;212&gt; PRT &lt;213&gt; Homo sapiens &lt;400 18

<td>Gin 1</td><td>Cheese</td><td>Pro</td><td>Cheese</td><td>Cheese 5</td><td>Leu</td><td>Cheese</td><td>Ala</td><td>Cheese</td><td>Val 10</td><td>Gly</td><td>Asp</td><td>Arg</td><td>Val</td><td>Thr 15</td><td>How much</td>

<td>Thr</td><td>Cys</td><td>Arg</td><td>Ala</td><td>Cheese</td><td>Gin</td><td>Cheese</td><td>How much</td><td>Asn</td><td>Thr</td><td>Tyr</td><td>Leu</td><td>How much</td><td>Trp</td><td>Tyr</td><td>Gin</td>

<td></td><td></td><td></td><td> 20</td><td></td><td></td><td></td><td></td><td> 25</td><td></td><td></td><td></td><td></td><td> 30</td><td></td><td></td>

<td>Gin</td><td>Lys</td><td>Pro</td><td>Gly</td><td>Lys</td><td>Ala</td><td>Pro</td><td>Asn</td><td>Phe</td><td>Leu</td><td>how much</td><td>Cheese</td><td>Ala</td><td>Thr</td><td>Cheese</td><td>How much</td>

<td></td><td></td><td> 35</td><td></td><td></td><td></td><td></td><td> 40</td><td></td><td></td><td></td><td></td><td> 45</td><td></td><td></td><td></td>

<td>Leu</td><td>Gin</td><td>Cheese</td><td>Gly</td><td>Val</td><td>Pro</td><td>Cheese</td><td>Arg</td><td>Phe</td><td>Arg</td><td>Gly</td><td>Cheese</td><td>dy</td><td>Cheese</td><td>Gly</td><td>Thr</td>

<td></td><td> 50</td><td></td><td></td><td></td><td></td><td> 55</td><td></td><td></td><td></td><td></td><td> 60</td><td></td><td></td><td></td><td></td>

<td>Asn</td><td>Phe</td><td>Thr</td><td>Leu</td><td>Thr</td><td>How much</td><td>Asn</td><td>Cheese</td><td>Leu</td><td>His</td><td>Pro</td><td>Glu</td><td>Asp</td><td>Phe</td><td>Ala</td><td>Thr</td>

<td> 65</td><td></td><td></td><td></td><td></td><td> 70</td><td></td><td></td><td></td><td></td><td> 75</td><td></td><td></td><td></td><td></td><td> 80</td>

<td>Tyr</td><td>Tyr</td><td>Cys</td><td>Gin</td><td>Gin</td><td>Cheese</td><td>Tyr</td><td>Cheese</td><td>Thr</td><td>Pro</td><td>Phe</td><td>Thr</td><td>Phe</td><td>Gly</td><td>Pro</td><td>Gly</td>

<td></td><td></td><td></td><td></td><td> 85</td><td></td><td></td><td></td><td></td><td> 90</td><td></td><td></td><td></td><td></td><td> 95</td><td></td>

<td>Thr</td><td>Lys</td><td>Val</td><td>Asp</td><td>How much</td><td>Lys</td><td>Arg</td><td>Thr</td><td>Val</td><td>Ala</td><td>Ala</td><td>Pro</td><td>Cheese</td><td>Val</td><td>Phe</td><td>How much</td>

<td></td><td></td><td></td><td> 100</td><td></td><td></td><td></td><td></td><td> 105</td><td></td><td></td><td></td><td></td><td> 110</td><td></td><td></td>

<td>Phe</td><td>Pro</td><td>Pro</td><td>Cheese</td><td>Asp</td><td>Glu</td><td>Gin</td><td>Leu</td><td>Lys</td><td>Cheese</td><td>Gly</td><td>Thr</td><td>Ala</td><td>Cheese</td><td>Val</td><td>Val</td>

<td></td><td></td><td> 115</td><td></td><td></td><td></td><td></td><td> 120</td><td></td><td></td><td></td><td></td><td> 125</td><td></td><td></td><td></td>

<td>Cys</td><td>Leu</td><td>Leu</td><td>Asn</td><td>Asn</td><td>Phe</td><td>Tyr</td><td>Pro</td><td>Arg</td><td>Glu</td><td>Al a</td><td>Lys</td><td>Val</td><td>Gin</td><td>Trp</td><td>Lys</td>

130 135 140

Val Asp Asn Ala Leu Gin Ser Gly

145 150

PL 214 003 Β1 &lt;210&gt; 19 &lt;211&gt; 142 &lt;212&gt; PRT &lt;213&gt; Homo sapiens &lt;400&gt; 19

<td>Cheese 1</td><td>Pro</td><td>Gly</td><td>Thr</td><td>Leu 5</td><td>Cheese</td><td>Leu</td><td>Cheese</td><td>Pro</td><td>Gly 10</td><td>Glu</td><td>Arg</td><td>Ala</td><td>Thr</td><td>Leu 15</td><td>Cheese</td>

<td>Cys</td><td>Arg</td><td>Ala</td><td>Cheese</td><td>Gin</td><td>Cheese</td><td>How much</td><td>Cheese</td><td>Cheese</td><td>Asn</td><td>Phe</td><td>Leu</td><td>Ala</td><td>Trp</td><td>Tyr</td><td>Gin</td>

<td></td><td></td><td></td><td> 20</td><td></td><td></td><td></td><td></td><td> 25</td><td></td><td></td><td></td><td></td><td> 30</td><td></td><td></td>

<td>Gin</td><td>Bald</td><td>Pro</td><td>dy</td><td>Gin</td><td>Ala</td><td>Pro</td><td>Arg</td><td>Leu</td><td>Leu</td><td>How much</td><td>Tyr</td><td>Arg</td><td>Pro</td><td>Cheese</td><td>Cheese</td>

<td></td><td></td><td> 35</td><td></td><td></td><td></td><td></td><td> 40</td><td></td><td></td><td></td><td></td><td> 45</td><td></td><td></td><td></td>

<td>Arg</td><td>Ala</td><td>Thr</td><td>Gly</td><td>How much</td><td>Pro</td><td>Asp</td><td>Cheese</td><td>Phe</td><td>Cheese</td><td>Gly</td><td>Cheese</td><td>Gly</td><td>Cheese</td><td>Gly</td><td>Thr</td>

<td></td><td> 50</td><td></td><td></td><td></td><td></td><td> 55</td><td></td><td></td><td></td><td></td><td> 60</td><td></td><td></td><td></td><td></td>

<td>Asp</td><td>Phe</td><td>Thr</td><td>Leu</td><td>Thr</td><td>How much</td><td>Cheese</td><td>Arg</td><td>Leu</td><td>Glu</td><td>Pro</td><td>Glu</td><td>Asp</td><td>Phe</td><td>Ala</td><td>Leu</td>

<td> 65</td><td></td><td></td><td></td><td></td><td> 70</td><td></td><td></td><td></td><td></td><td> 75</td><td></td><td></td><td></td><td></td><td>ao</td>

<td>Tyr</td><td>Tyr</td><td>Cys</td><td>Gin</td><td>Gin</td><td>Tyr</td><td>Gly</td><td>Thr</td><td>Cheese</td><td>Pro</td><td>Phe</td><td>Thr</td><td>Phe</td><td>Gly</td><td>Pro</td><td>Gly</td>

<td></td><td></td><td></td><td></td><td> 85</td><td></td><td></td><td></td><td></td><td> 90</td><td></td><td></td><td></td><td></td><td> 95</td><td></td>

<td>Thr</td><td>Lys</td><td>vai</td><td>Asp</td><td>How much</td><td>Bald</td><td>Arg</td><td>Thr</td><td>Val</td><td>Ala</td><td>Ala</td><td>Pro</td><td>Cheese</td><td>Val</td><td>Phe</td><td>How much</td>

<td></td><td></td><td></td><td> 100</td><td></td><td></td><td></td><td></td><td> 105</td><td></td><td></td><td></td><td></td><td> 110</td><td></td><td></td>

<td>Phe</td><td>Pro</td><td>Pro</td><td>Cheese</td><td>Asp</td><td>Glu</td><td>Gin</td><td>Leu</td><td>Lys</td><td>Cheese</td><td>Gly</td><td>Thr</td><td>Ala</td><td>Cheese</td><td>val</td><td>val</td>

<td></td><td></td><td> 115</td><td></td><td></td><td></td><td></td><td> 120</td><td></td><td></td><td></td><td></td><td> 125</td><td></td><td></td><td></td>

<td>Cys</td><td>Leu</td><td>Leu</td><td>Asn</td><td>Asn</td><td>Phe</td><td>Tyr</td><td>Pro</td><td>Arg</td><td>Glu</td><td>Ala</td><td>Lys</td><td>Val</td><td>Gin</td><td></td><td></td>

130 135 140 &lt;210&gt; 20 &lt;211&gt; 155 &lt;212&gt; PRT &lt;213&gt; Homo sapiens

<td colspan="5"> &lt;400&gt; 20</td><td rowspan="2">Cheese</td><td rowspan="2">Val</td><td rowspan="2">Thr</td>

<td>Cheese 1</td><td>Pro</td><td>Asp</td><td>Phe</td><td>Gin 5</td>

<td>Cys</td><td>Arg</td><td>Ala</td><td>Cheese twenty</td><td>G1 n</td><td>Cheese</td><td>How much</td><td>Gly</td>

<td>Lys</td><td>Pro</td><td>Asp 35</td><td>Gin</td><td>Cheese</td><td>Pro</td><td>Lys</td><td>Leu 40</td>

<td>Phe</td><td>Cheese 50</td><td>Gx y</td><td>Val</td><td>Pro</td><td>Cheese</td><td>Arg 55</td><td>Phe</td>

<td>Phe 65</td><td>Thr</td><td>Leu</td><td>Thr</td><td>How much</td><td>Asn 70</td><td>Cheese</td><td>Leu</td>

<td>Tyr</td><td>Cys</td><td>His</td><td>Gin</td><td>Cheese 85</td><td>Cheese</td><td>Cheese</td><td>Leu</td>

<td>Pro</td><td>Lys 10</td><td>Glu</td><td>Lys</td><td>Val</td><td>Thr</td><td>how much 15</td><td>Thr</td>

<td>Cheese 25</td><td>Cheese</td><td>Leu</td><td>His</td><td>Trp</td><td>Tyr thirty</td><td>Gin.</td><td>Gin</td>

<td>Leu</td><td>How much</td><td>Lys</td><td>Tyr</td><td>Ala 45</td><td>Cheese</td><td>Gin</td><td>Cheese</td>

<td>Cheese</td><td>Gly</td><td>Cheese</td><td>Gly 60</td><td>Cheese</td><td>Gly</td><td>Thr</td><td>Asp</td>

<td>Glu</td><td>Ala</td><td>Glu 75</td><td>Asp</td><td>Ala</td><td>Ala</td><td>Thr</td><td>Tyr 80</td>

<td>Pro</td><td>Leu 90</td><td>Thr</td><td>Phe</td><td>Gly</td><td>Gly</td><td>Gly 95</td><td>Thr</td>

PL 214 003 Β1

<td>Lys</td><td>Val</td><td>1 u</td><td>And ..i. Θ 100</td><td>Lys</td><td>Arg</td><td>Thr</td><td>Val</td><td>Ala 105</td><td>Ala</td><td>Pro</td><td>Cheese</td><td>Val</td><td>Phe 110</td><td>How much</td><td>Phe</td>

<td>Pro</td><td>Pro</td><td>Cheese</td><td>Asp</td><td>Glu</td><td>Gin</td><td>Leu</td><td>Lys</td><td>Cheese</td><td>Gly</td><td>Thr</td><td>Ala</td><td>Cheese</td><td>Val</td><td>Val</td><td>Cys</td>

<td></td><td></td><td> 115</td><td></td><td></td><td></td><td></td><td> 120</td><td></td><td></td><td></td><td></td><td> 125</td><td></td><td></td><td></td>

<td>Leu</td><td>Leu</td><td>Asn</td><td>Asn</td><td>Phe</td><td>Tyr</td><td>Pro</td><td>Arg</td><td>Glu</td><td>Ala</td><td>Lys</td><td>Val</td><td>Gin</td><td>Trp</td><td>Lys</td><td>Val</td>

<td></td><td> 130</td><td></td><td></td><td></td><td></td><td> 135</td><td></td><td></td><td></td><td></td><td> 140</td><td></td><td></td><td></td><td></td>

<td>Asp</td><td>Asn</td><td>Ala</td><td>Leu</td><td>Gin</td><td>Cheese</td><td>Gly</td><td>Asn</td><td>Cheese</td><td>Gin</td><td>Glu</td><td></td><td></td><td></td><td></td><td></td>

145 150 155 &lt;210&gt; 21 &lt;211&gt; 146 &lt;212&gt; PRT

<td colspan="3">&lt;213&gt; Homo</td><td colspan="13">sapiens</td>

<td colspan="3"> &lt;400&gt; 21</td><td></td><td></td><td></td><td></td><td></td><td></td><td></td><td></td><td></td><td></td><td></td><td></td><td></td>

<td>Gin</td><td>Cheese</td><td>Pro</td><td>Gly</td><td>Thr</td><td>i_iSU</td><td>Cheese</td><td>Leu</td><td>Cheese</td><td>Pro</td><td>Gly</td><td>Glu</td><td>Ar g</td><td>Al a</td><td> ^|**·</td><td>L ^ su</td>

<td> 1.</td><td></td><td></td><td></td><td> 5</td><td></td><td></td><td></td><td></td><td> 10</td><td></td><td></td><td></td><td></td><td> 15</td><td></td>

<td>Cheese</td><td>cys</td><td>Arg</td><td>Ala</td><td>Cheese</td><td>Gin</td><td>Cheese</td><td>val</td><td>Cheese</td><td>Cheese</td><td>Tyr</td><td>Leu</td><td>Ala</td><td>Trp</td><td>Tyr</td><td>Gin</td>

<td></td><td></td><td></td><td> 20</td><td></td><td></td><td></td><td></td><td> 25</td><td></td><td></td><td></td><td></td><td> 30</td><td></td><td></td>

<td>Gin</td><td>Lys</td><td>Pro</td><td>Gly</td><td>Gin</td><td>Ala</td><td>Pro</td><td>Arg</td><td>Leu</td><td>Leu</td><td>How much</td><td>Tyr</td><td>Gly</td><td>Ala</td><td>Cheese</td><td>Cheese</td>

<td></td><td></td><td> 35</td><td></td><td></td><td></td><td></td><td> 40</td><td></td><td></td><td></td><td></td><td> 45</td><td></td><td></td><td></td>

<td>Arg</td><td>Ala</td><td>Thr</td><td>Gly</td><td>How much</td><td>Pro</td><td>Asp</td><td>Arg</td><td>Phe</td><td>Cheese</td><td>Gly</td><td>Cheese</td><td>Gly</td><td>Cheese</td><td>Gly</td><td>Thr</td>

<td></td><td> 50</td><td></td><td></td><td></td><td></td><td> 55</td><td></td><td></td><td></td><td></td><td> 60</td><td></td><td></td><td></td><td></td>

<td>Asp</td><td>Phe</td><td>Thr</td><td>Leu</td><td>Thr</td><td>How much</td><td>Cheese</td><td>Arg</td><td>Leu</td><td>Glu</td><td>Pro</td><td>Glu</td><td>Asp</td><td>Phe</td><td>Ala</td><td>Val</td>

<td> 65</td><td></td><td></td><td></td><td></td><td> 70</td><td></td><td></td><td></td><td></td><td> 75</td><td></td><td></td><td></td><td></td><td> 80</td>

<td>Tyr</td><td>Tyr</td><td>Cys</td><td>Gin</td><td>Gin</td><td>Tyr</td><td>Gly</td><td>Arg</td><td>Cheese</td><td>Pro</td><td>Phe</td><td>Thr</td><td>Phe</td><td>Gly</td><td>Pro</td><td>Gly</td>

<td></td><td></td><td></td><td></td><td>B5</td><td></td><td></td><td></td><td></td><td> 90</td><td></td><td></td><td></td><td></td><td> 95</td><td></td>

<td>Thr</td><td>Lys</td><td>val</td><td>Asp</td><td>How much</td><td>Lys</td><td>Arg</td><td>T m</td><td>Val</td><td>Ala</td><td>Ala</td><td>Pro</td><td>Cheese</td><td>Val</td><td>Phe</td><td>How much</td>

<td></td><td></td><td></td><td> 100</td><td></td><td></td><td></td><td></td><td> 105</td><td></td><td></td><td></td><td></td><td> 110</td><td></td><td></td>

<td>Phe</td><td>Pro</td><td>Pro</td><td>Cheese</td><td>Asp</td><td>G Lu</td><td>Gin</td><td>Leu</td><td>Lys</td><td>Cheese</td><td>Gly</td><td>Thr</td><td>Ala</td><td>Cheese</td><td>Val</td><td>Val</td>

<td></td><td></td><td> 115</td><td></td><td></td><td></td><td></td><td> 120</td><td></td><td></td><td></td><td></td><td> 125</td><td></td><td></td><td></td>

<td>Cys</td><td>Leu</td><td>Leu</td><td>Asn</td><td>Asn</td><td>Phe</td><td>Tyr</td><td>Pro</td><td>Arg</td><td>Glu</td><td>Ala</td><td>Lys</td><td>Val</td><td>Gin</td><td>Trp</td><td>Lys</td>

130 135 140

Gly Gly

145 &lt;210&gt; 22 &lt;211&gt; 139 &lt;212&gt; PRT &lt;213&gt; Homo sapiens &lt;400&gt; 22

Pro Ser Ser Leu Ser Ala Ser Val Gly Asp Arg Val Thr Ile Thr Cys

10 15

PL 214 003 Β1

<td>Arg</td><td>Ala</td><td>Cheese</td><td>Gin twenty</td><td>Cheese</td><td>How much</td><td>Asn</td><td>Cheese</td><td>Tyr 25</td><td>Leu</td><td>Asp</td><td>Trp</td><td>Tyr</td><td>Gin thirty</td><td>Gin</td><td>Lys</td>

<td>Pro</td><td>Gly</td><td>Lys</td><td>Ala</td><td>Pro</td><td>Lys</td><td>Leu</td><td>Leu</td><td>How much</td><td>Tyr</td><td>Ala</td><td>Ala</td><td>Cheese</td><td>Cheese</td><td>Leu</td><td>Gin</td>

<td></td><td></td><td> 35</td><td></td><td></td><td></td><td></td><td> 40</td><td></td><td></td><td></td><td></td><td> 45</td><td></td><td></td><td></td>

<td>Cheese</td><td>Gly</td><td>Val</td><td>Pro</td><td>Cheese</td><td>Arg</td><td>Phe</td><td>Cheese</td><td>Gly</td><td>Cheese</td><td>Gly</td><td>Cheese</td><td>Gly</td><td>Thr</td><td>Asp</td><td>Phe</td>

<td></td><td> 50</td><td></td><td></td><td></td><td></td><td> 55</td><td></td><td></td><td></td><td></td><td> 60</td><td></td><td></td><td></td><td></td>

<td>Thr</td><td>Leu</td><td>Thr</td><td>How much</td><td>Cheese</td><td>Cheese</td><td>Leu</td><td>Gin</td><td>Pro</td><td>Glu</td><td>Asp</td><td>Phe</td><td>Ala</td><td>Thr</td><td>Tyr</td><td>Tyr</td>

<td> 65</td><td></td><td></td><td></td><td></td><td> 70</td><td></td><td></td><td></td><td></td><td> 75</td><td></td><td></td><td></td><td></td><td> 80</td>

<td>Cys</td><td>Gin</td><td>Gin</td><td>Tyr</td><td>Tyr</td><td>Cheese</td><td>Thr</td><td>Pro</td><td>Phe</td><td>Thr</td><td>Phe</td><td>Gly</td><td>Pro</td><td>Gly</td><td>Thr</td><td>Lys</td>

<td></td><td></td><td></td><td></td><td> 85</td><td></td><td></td><td></td><td></td><td> 90</td><td></td><td></td><td></td><td></td><td> 95</td><td></td>

<td>Val</td><td>Glu</td><td>How much</td><td>Lys</td><td>Arg</td><td>Thr</td><td>Val</td><td>Ala</td><td>Al a</td><td>Pro</td><td>Cheese</td><td>Val</td><td>Phe</td><td>How much</td><td>Phe</td><td>Pro</td>

<td></td><td></td><td></td><td> 100</td><td></td><td></td><td></td><td></td><td> 105</td><td></td><td></td><td></td><td></td><td> 110</td><td></td><td></td>

<td>Pro</td><td>Cheese</td><td>Asp</td><td>Glu</td><td>Gin</td><td>Leu</td><td>Lys</td><td>Cheese</td><td>Gly</td><td>Thr</td><td>Ala</td><td>Cheese</td><td>Val</td><td>Val</td><td>Cys</td><td>Leu</td>

<td></td><td></td><td> 115</td><td></td><td></td><td></td><td></td><td> 120</td><td></td><td></td><td></td><td></td><td> 125</td><td></td><td></td><td></td>

<td>Leu</td><td>Asn</td><td>Asn</td><td>Phe</td><td>Tyr</td><td>Pro</td><td>Arg</td><td>Glu</td><td>Ala</td><td>Lys</td><td>Val</td><td></td><td></td><td></td><td></td><td></td>

130 135 &lt;210&gt; 23 &lt;211&gt; 134 &lt;212&gt; PRT &lt;213&gt; Homo sapiens &lt;400&gt; 23

<td>Thr 1</td><td>Gin</td><td>Cheese</td><td>Pro</td><td>Cheese 5</td><td>Cheese</td><td>Leu</td><td>Cheese</td><td>Ala</td><td>Cheese 10</td><td>Val</td><td>Gly</td><td>Asp</td><td>Arg</td><td>Val 15</td><td>Thr</td>

<td>How much</td><td>Thr</td><td>Cys</td><td>Arg</td><td>Ala</td><td>Cheese</td><td>Gin</td><td>Asn</td><td>How much</td><td>Cheese</td><td>Arg</td><td>Tyr</td><td>Leu</td><td>Asn</td><td>Trp</td><td>Tyr</td>

<td></td><td></td><td></td><td> 20</td><td></td><td></td><td></td><td></td><td> 25</td><td></td><td></td><td></td><td></td><td> 30</td><td></td><td></td>

<td>G1 n</td><td>Gin</td><td>Lys</td><td>Pro</td><td>Gly</td><td>Lys</td><td>Ala</td><td>Pro</td><td>Lys</td><td>Phe</td><td>Leu</td><td>How much</td><td>Tyr</td><td>Val</td><td>Ala</td><td>Cheese</td>

<td></td><td></td><td> 35</td><td></td><td></td><td></td><td></td><td> 40</td><td></td><td></td><td></td><td></td><td> 45</td><td></td><td></td><td></td>

<td>How much</td><td>Leu</td><td>Gin</td><td>Cheese</td><td>Gly</td><td>Val</td><td>Pro</td><td>Cheese</td><td>Gly</td><td>Phe</td><td>Cheese</td><td>Ala</td><td>Cheese</td><td>Gly</td><td>Cheese</td><td>Gly</td>

<td></td><td> 50</td><td></td><td></td><td></td><td></td><td> 55</td><td></td><td></td><td></td><td></td><td> 60</td><td></td><td></td><td></td><td></td>

<td>Pro</td><td>Asp</td><td>Phe</td><td>Thr</td><td>Leu</td><td>Thr</td><td>How much</td><td>Cheese</td><td>Cheese</td><td>Leu</td><td>Gin</td><td>Pro</td><td>Glu</td><td>Asp</td><td>Phe</td><td>Ala</td>

<td> 65</td><td></td><td></td><td></td><td></td><td> 70</td><td></td><td></td><td></td><td></td><td> 75</td><td></td><td></td><td></td><td></td><td> 80</td>

<td>Thr</td><td>Tyr</td><td>Tyr</td><td>Cys</td><td>Gin</td><td>Gin</td><td>Cheese</td><td>Tyr</td><td>Cheese</td><td>Thr</td><td>Pro</td><td>Phe</td><td>Thr</td><td>Phe</td><td>Gly</td><td>Pro</td>

<td></td><td></td><td></td><td></td><td> 85</td><td></td><td></td><td></td><td></td><td> 90</td><td></td><td></td><td></td><td></td><td> 95</td><td></td>

<td>Gly</td><td>Thr</td><td>Lys</td><td>Val</td><td>Asp</td><td>How much</td><td>Lys</td><td>Arg</td><td>Thr</td><td>Val</td><td>Ala</td><td>Ala</td><td>Pro</td><td>Cheese</td><td>Val</td><td>Phe</td>

<td></td><td></td><td></td><td> 100</td><td></td><td></td><td></td><td></td><td> 105</td><td></td><td></td><td></td><td></td><td> 110</td><td></td><td></td>

<td>How much</td><td>Phe</td><td>Pro</td><td>Pro</td><td>Cheese</td><td>Asp</td><td>Glu</td><td>Gin</td><td>Leu</td><td>Lys</td><td>Cheese</td><td>Gly</td><td>Thr</td><td>Ala</td><td>Cheese</td><td>Val</td>

<td></td><td></td><td> 115</td><td></td><td></td><td></td><td></td><td> 120</td><td></td><td></td><td></td><td></td><td> 125</td><td></td><td></td><td></td>

Val Cys Leu Leu Asn Asn

130

PL 214 003 Β1 &lt;210&gt; 24 &lt;211&gt; 150 &lt;212&gt; PRT &lt;213&gt; Homo sapiens &lt;400&gt; 24

<td>Thr 1</td><td>Gin</td><td>Cheese</td><td colspan="2">Pro Ser 5</td><td>Cheese</td><td>Leu</td><td>Cheese</td><td>Ala</td><td>Cheese 10</td><td>Val</td><td>dy</td><td>Asp</td><td>Arg</td><td>Val 15</td><td>Thr</td>

<td>How much</td><td>Thr</td><td>Cys</td><td>Arg</td><td>Ala</td><td>Cheese</td><td>Gin</td><td>Cheese</td><td>How much</td><td>Cys</td><td>Asn</td><td>Tyr</td><td>Leu</td><td>Asn</td><td>Trp</td><td>Tyr</td>

<td></td><td></td><td></td><td> 20</td><td></td><td></td><td></td><td></td><td> 2</td><td></td><td></td><td></td><td></td><td> 30</td><td></td><td></td>

<td>Gin</td><td>Gin</td><td>Lys</td><td>Pro</td><td>Gly</td><td>Lys</td><td>Ala</td><td>Pro</td><td>Arg</td><td>Val</td><td>Leu</td><td>How much</td><td>Tyr</td><td>Ala</td><td>Ala</td><td>Cheese</td>

<td></td><td></td><td> 35</td><td></td><td></td><td></td><td></td><td> 40</td><td></td><td></td><td></td><td></td><td> 45</td><td></td><td></td><td></td>

<td>Cheese</td><td>Leu</td><td>Gin</td><td>Gly</td><td>Gly</td><td>Val</td><td>Pro</td><td>Cheese</td><td>Arg</td><td>Phe</td><td>Cheese</td><td>Gly</td><td>Cheese</td><td>Giy</td><td>Cheese</td><td>Gly</td>

<td></td><td> 50</td><td></td><td></td><td></td><td></td><td> 55</td><td></td><td></td><td></td><td></td><td> 60</td><td></td><td></td><td></td><td></td>

<td>How much</td><td>Asp</td><td>Cys</td><td>Thr</td><td>Leu</td><td>Thr</td><td>How much</td><td>Cheese</td><td>Cheese</td><td>Leu</td><td>Gin</td><td>Pro</td><td>Glu</td><td>Asp</td><td>phe</td><td>Ala</td>

<td> 65</td><td></td><td></td><td></td><td></td><td> 70</td><td></td><td></td><td></td><td></td><td> 75</td><td></td><td></td><td></td><td></td><td> 80</td>

<td>Thr</td><td>Tyr</td><td>Tyr</td><td>Cys</td><td>Gin</td><td>Gin</td><td>Cheese</td><td>Tyr</td><td>How much</td><td>Thr</td><td>Pro</td><td>Phe</td><td>Thr</td><td>Phe</td><td>Gly</td><td>Pro</td>

<td></td><td></td><td></td><td></td><td> 85</td><td></td><td></td><td></td><td></td><td> 90</td><td></td><td></td><td></td><td></td><td> 95</td><td></td>

<td>Gly</td><td>Thr</td><td>Arg</td><td>Val</td><td>Asp</td><td>How much</td><td>Ir U</td><td>Arg</td><td>Thr</td><td>Val</td><td>Ala</td><td>Ala</td><td>Pro</td><td>Cheese</td><td>Val</td><td>Phe</td>

<td></td><td></td><td></td><td> 100</td><td></td><td></td><td></td><td></td><td> 105</td><td></td><td></td><td></td><td></td><td> 110</td><td></td><td></td>

<td>How much</td><td>Phe</td><td>Pro</td><td>Pro</td><td>Cheese</td><td>Asp</td><td>Glu</td><td>Gin</td><td>Leu</td><td>Lys</td><td>Cheese</td><td>Gly</td><td>Thr</td><td>Ala</td><td>Cheese</td><td>Val</td>

<td></td><td></td><td> 115</td><td></td><td></td><td></td><td></td><td> 120</td><td></td><td></td><td></td><td></td><td> 125</td><td></td><td></td><td></td>

<td>Va 1</td><td>Cys</td><td>Leu</td><td>Leu</td><td>Asn</td><td>Asn</td><td>Phe</td><td>Tyr</td><td>Pro</td><td>Ar g</td><td>Glu</td><td>Ala</td><td>Lys</td><td>val</td><td>Gin</td><td>Trp</td>

<td></td><td> 130</td><td></td><td></td><td></td><td></td><td> 135</td><td></td><td></td><td></td><td></td><td> 140</td><td></td><td></td><td></td><td></td>

<td>Lys</td><td>Val</td><td>Asp</td><td>Asn</td><td>Ala</td><td>Tyr</td><td></td><td></td><td></td><td></td><td></td><td></td><td></td><td></td><td></td><td></td>

145 150 &lt;210&gt; 25 &lt;211&gt; 139 &lt;212&gt; PRT &lt;213&gt; Homo sapiens &lt;400&gt; 25

<td>Pro 1</td><td>leu</td><td>Cheese</td><td>Leu</td><td colspan="2">Pro Val 5</td><td>Thr</td><td colspan="2">Leu Gly</td><td>Gin 10</td><td>Pro</td><td>Ala</td><td>Cheese</td><td>How much</td><td>Cheese 15</td><td>Cys</td>

<td>Arg</td><td>Cheese</td><td>Cheese</td><td>Gin</td><td>Cheese</td><td>Leu</td><td>Val</td><td>Tyr</td><td>Cheese</td><td>Asp</td><td>Gly</td><td>Asn</td><td>Thr</td><td>Tyr</td><td>Leu</td><td>Asn</td>

<td></td><td></td><td></td><td> 20</td><td></td><td></td><td></td><td></td><td> 25</td><td></td><td></td><td></td><td></td><td> 30</td><td></td><td></td>

<td>Trp</td><td>Phe</td><td>Gin</td><td>Gin</td><td>Arg</td><td>Pro</td><td>Gly</td><td>Gin</td><td>Cheese</td><td>Pro</td><td>Arg</td><td>Arg</td><td>Leu</td><td>How much</td><td>Tyr</td><td>Lys</td>

<td></td><td></td><td> 35</td><td></td><td></td><td></td><td></td><td> 40</td><td></td><td></td><td></td><td></td><td> 45</td><td></td><td></td><td></td>

<td>Val</td><td>Cheese</td><td>Asn</td><td>Trp</td><td>Asp</td><td>Cheese</td><td>Giy</td><td>Val</td><td>Pro</td><td>Asp</td><td>Arg</td><td>Phe</td><td>Cheese</td><td>Gly</td><td>Cheese</td><td>Gly</td>

<td></td><td> 50</td><td></td><td></td><td></td><td></td><td> 55</td><td></td><td></td><td></td><td></td><td> 60</td><td></td><td></td><td></td><td></td>

Ser Gly Thr Asp Phe Thr Leu Lys Ile Ser Arg Val Glu Ala Glu Asp

PL 214 003 Β1

<td colspan="3"> 65</td><td colspan="4"> 70</td><td colspan="5"> 75</td><td colspan="4"> 80</td>

<td>Val</td><td>Gly</td><td>Val</td><td>Tyr</td><td>Tyr</td><td>Cys</td><td>Underworld</td><td>Gin</td><td>Gly</td><td>Cheese</td><td>His</td><td>Trp</td><td>Pro</td><td>Pro</td><td>Thr</td><td>Phe</td>

<td></td><td></td><td></td><td></td><td> 85</td><td></td><td></td><td></td><td></td><td> 90</td><td></td><td></td><td></td><td></td><td> 95</td><td></td>

<td>Gly</td><td>Gin</td><td>Gly</td><td>Thr</td><td>Lys</td><td>Val</td><td>Glu</td><td>How much</td><td>Lys</td><td>Arg</td><td>Thr</td><td>Val</td><td>Ala</td><td>Ala</td><td>Pro</td><td>Cheese</td>

<td></td><td></td><td></td><td> 100</td><td></td><td></td><td></td><td></td><td> 105</td><td></td><td></td><td></td><td></td><td> 110</td><td></td><td></td>

<td>Val</td><td>Phe</td><td>how much</td><td>Phe</td><td>Pro</td><td>Pro</td><td>Cheese</td><td>Asp</td><td>Glu</td><td>Gin</td><td>Leu</td><td>Lys</td><td>Cheese</td><td>Gly</td><td>Thr</td><td>Ala</td>

<td></td><td></td><td> 115</td><td></td><td></td><td></td><td></td><td> 120</td><td></td><td></td><td></td><td></td><td> 125</td><td></td><td></td><td></td>

<td>Cheese</td><td>Val</td><td>Val</td><td>Cys</td><td>Leu</td><td>Leu</td><td>Asn</td><td>Asn</td><td>Phe</td><td>Tyr</td><td>Pro</td><td></td><td></td><td></td><td></td><td></td>

130 135 &lt;210&gt; 26 &lt;211&gt; 133 &lt;212&gt; PRT

<td colspan="3">&lt;213&gt; Homo</td><td colspan="13">sapiens</td>

<td colspan="3"> &lt;400&gt; 26</td><td></td><td></td><td></td><td></td><td></td><td></td><td></td><td></td><td></td><td></td><td></td><td></td><td></td>

<td>Pro</td><td>Gly</td><td>Glu</td><td>Pro</td><td>Al a</td><td>Cheese</td><td>How much</td><td>Cheese</td><td>Cys</td><td>Arg</td><td>Cheese</td><td>Cheese</td><td>Gin</td><td>Cheese</td><td>Leu</td><td>Leu</td>

<td> 1</td><td></td><td></td><td></td><td> 5</td><td></td><td></td><td></td><td></td><td> 10</td><td></td><td></td><td></td><td></td><td> 15</td><td></td>

<td>His</td><td>Cheese</td><td>Asn</td><td>Gly</td><td>Tyr</td><td>Asn</td><td>Tyr</td><td>Leu</td><td>Asp</td><td>Trp</td><td>Tyr</td><td>Leu</td><td>Gin</td><td>Lys</td><td>Pro</td><td>Gly</td>

<td></td><td></td><td></td><td> 20</td><td></td><td></td><td></td><td></td><td> 25</td><td></td><td></td><td></td><td></td><td> 30</td><td></td><td></td>

<td>Gin</td><td>Cheese</td><td>Pro</td><td>Gin</td><td>Leu</td><td>Leu</td><td>How much</td><td>Tyr</td><td>Leu</td><td>Gly</td><td>Cheese</td><td>Asn</td><td>Arg</td><td>Ala</td><td>Cheese</td><td>Gly</td>

<td></td><td></td><td> 35</td><td></td><td></td><td></td><td></td><td> 40</td><td></td><td></td><td></td><td></td><td> 45</td><td></td><td></td><td></td>

<td>Val</td><td>Pro</td><td>Asp</td><td>Arg</td><td>Phe</td><td>Cheese</td><td>Gly</td><td>Cheese</td><td>Gly</td><td>Cheese</td><td>Gly</td><td>Thr</td><td>Asp</td><td>Phe</td><td>Thr</td><td>Leu</td>

<td></td><td> 50</td><td></td><td></td><td></td><td></td><td> 55</td><td></td><td></td><td></td><td></td><td> 60</td><td></td><td></td><td></td><td></td>

<td>Lys</td><td>Leu</td><td>Cheese</td><td>Arg</td><td>Val</td><td>Glu</td><td>Ala</td><td>Glu</td><td>Asp</td><td>Val</td><td>Gly</td><td>Val</td><td>Tyr</td><td>Tyr</td><td>Cys</td><td>Underworld</td>

<td> 65</td><td></td><td></td><td></td><td></td><td> 70</td><td></td><td></td><td></td><td></td><td> 75</td><td></td><td></td><td></td><td></td><td> 80</td>

<td>Gin</td><td>Ala</td><td>Leu</td><td>Gin</td><td>Thr</td><td>Pro</td><td>Leu</td><td>Thr</td><td>Phe</td><td>Gly</td><td>Gly</td><td>Gly</td><td>Thr</td><td>Lys</td><td>val</td><td>Glu</td>

<td></td><td></td><td></td><td></td><td> 85</td><td></td><td></td><td></td><td></td><td> 90</td><td></td><td></td><td></td><td></td><td> 95</td><td></td>

<td>How much</td><td>Ly p</td><td>Arg</td><td>Thr</td><td>Vai</td><td>Al a</td><td>Al a</td><td>Pro</td><td>Cheese</td><td>ci. '. 1.</td><td>Phe</td><td>How much</td><td>Phe</td><td>Pro</td><td>Pro</td><td>Cheese</td>

<td></td><td></td><td></td><td> 100</td><td></td><td></td><td></td><td></td><td> 105</td><td></td><td></td><td></td><td></td><td> 110</td><td></td><td></td>

<td>Asp</td><td>Glu</td><td>Gin</td><td>Leu</td><td>Lys</td><td>Cheese</td><td>Gly</td><td>Thr</td><td>Ala</td><td>Cheese</td><td>Val</td><td>Val</td><td>Cys</td><td>Leu</td><td>Leu</td><td>Asn</td>

115 1.20 125

Asn Phe Tyr Pro Arg

130 &lt;210&gt; 27 &lt;211&gt; 1392 &lt;212&gt; DNA &lt;213&gt; Homo sapiens &lt;400&gt; 27 atggagtttg ggctgagctg ggttttcctc gttgctcttt taagaggtgt ccagtgtcag60 gtgcagctgg tggagtctgg gggaggcgtg gtccagcctg ggaggtccct gagactctcc120 cgtgtagcgt ctggattcac cttcagtagc catggcatgc actgggtccg ccaggctcca180

EN 214 003 Β1 ggcaaggggc gactccgtga caaatgaaca ttcggtcctt aagggcccat gccctgggct ggcgctctga tccctcagca aacgtagatc gtcgagtgcc ccaaaaccca gacgtgagcc cataatgcca gtcctcaccg aacaaaggcc gaaccacagg ctgacctgcc gggcagccgg ttcctctaca tgctccgtga ccgggtaaat tggagtgggt agggccgatt gcctgagagc ttgactactg cggtcttccc gcctggtcaa ccagcggcgt gcgtggtgac acaagcccag caccgtgccc aggacaccct acgaagaccc agacaaagcc ttgtgcacca tcccagcccc tgtacaccct tggtcaaagg agaacaacta gcaagctcac tgcatgaggc ga ggcagttata caccatctcc cgaggacacg gggccaggga cctggcgccc ggactacttc gcacaccttc cgtgccctcc caacaccaag agcaccacct catgatctcc cgaggtccag acgggaggag ggactggctg catcgagaaa gcccccatcc cttctacccc caagaccaca cgtggacaag tctgcacaac tggtatgatg agagacaatt gctgtgtatt accctggtca tgctccagga cccgaaccgg ccagctgtcc agcaacttcg gtggacaaga gtggcaggac cggacccctg ttcaactggt cagttcaaca aacggcaagg accatctcca cgggaggaga agcgacatcg cctcccatgc agcaggtggc cactacacgc gaagaaataa ccaagaacac actgtgcgag ccgtctcctc gcacctccga tgacggtgtc tacagtcctc gcacccagac cagttgagcg cgtcagtctt aggtcacgtg acgtggacgg gcacgttccg agtacaagtg aaaccaaagg tgaccaagaa ccgtggagtg tggactccga agcaggggaa agaagagcct atactatgca gctgtttctg aggaggtcac agcctccacc gagcacagcg gtggaactca aggactctac ctacacctgc caaatgttgt cctcttcccc cgtggtggtg cgtggaggtg tgtggtcagc caaggtctcc gcagccccga ccaggtcagc ggagagcaat cggctccttc cgtcttctca ctccctgtct

240

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1020

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1140

1200

1260

1320

1380

1392 &lt;210&gt; 28 &lt;211&gt; 1395 &lt;212&gt; DNA &lt;213&gt; Homo sapiens &lt;400&gt; 28 atggagtttg gtgcagctgg tgtacagcgt ggcaaggggc gactccgtga caaatgaaca ctggggtcct accaagggcc gcggccctgg tcaggcgctc tactccctca tgcaacgtag tgtgtcgagt cccccaaaac gtggacgtga gtgcataatg agcgtcctca tccaacaaag cgagaaccac agcctgacct aatgggcagc ttcttcctct tcatgctccg tctccgggta ggctgagctg tggagtctgg ctggattcac tggagtgggt agggccgatt gcctgagagc actttgacta catcggtctt gctgcctggt tgaccagcgg gcagcgtggt atcacaagcc gcccaccgtg ccaaggacac gccacgaaga ccaagacaaa ccgttgtgca gcctcccagc aggtgtacac gcctggtcaa cggagaacaa acagcaagct tgatgcatga aatga ggttttcctc gggaggcgtg cttcagtaac ggcagttata caccatctcc cgaggacacg ctggggccag ccccctggcg caaggactac cgtgcacacc gaccgtgccc cagcaacacc cccagcacca cctęatgatc ccccgaggtc gccacgggag ccaggactgg ccccatcgag cctgccccca aggcttctac ctacaagacc caccgtggac ggctctgcac gttgctcttt gtccagcctg tatggcatgc tggtatgatg agtgacaatt gctgtgtatt ggaaccctgg ccctgctcca ttccccgaac ttcccagctg tccagcaact aaggtggaca cctgtggcag tcccggaccc cagttcaact gagcagttca ctgaacggca aaaaccatct tcccgggagg cccagcgaca acacctccca aagagcaggt aaccactaca taagaggtgt ggaggtccct actgggtccg gaagtaataa ccaagaacac actgtgcgag tcaccgtctc ggagcacctc cggtgacggt tcctacagtc tcggcaccca agacagttga gaccgtcagt ctgaggtcac ggtacgtgga acagcacgtt aggagtacaa ccaaaaccaa agatgaccaa tcgccgtgga tgctggactc ggcagcaggg cgcagaagag ccagtgtcag gagact.ctcc ccaggctcca acactatgga gctgtatctg aggagagaga ctcagcctcc cgagagcaca gtcgtggaac ctcaggactc gacctacacc gcgcaaatgt cttcctcttc gtgcgtggtg cggcgtggag ccgtgtggtc gtgcaaggtc agggcagccc gaaccaggtc gtgggagagc cgacggctcc gaacgtcttc cctctccctg

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780

840

900

960

1020

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1140

1200

1260

1320

1380

1395 &lt;210&gt; 29 &lt;211&gt; 439 &lt;212&gt; DNA

GB 214 003 Β1 &lt;213&gt; Homo sapiens &lt;400&gt; 29 cctgggaggt atccactggg gatggaagaa aattccaaga tattactgtg gtcaccgtct aggagcacct ccggtgacgg gtcctacag ccctgagact tccgccaggc ataaagacta agacgctgta cgagagtggc cctcagcctc ccgagagcac tgtcgtggaa ctcctgtgca tccaggcaag tgcagactcc tttgcaaatg cccactgggg caccaagggc agcggccctg ctcaggcgct gcgtctggat gggctggagt gtgaagggcc aacagcctga ccacttgact ccatcggtct ggctgcctgg ctgaccagcg tcaccttcag gggtggcagt gattcaccat gagccgagga actggggcca tccccctggc tcaaggacta gcgtgcacac tagtcatggc tatatggtat ctccagagac cacggctgtg gggaaccctg gccctgctcc cttccccgaa cttcccagct

120

180

240

300

360

420

480

489 &lt;210&gt; 30 &lt;211&gt; 1392 &lt;212&gt; DNA &lt;213&gt; Homo sapiens &lt;400&gt; 30 atggagtttg gtgcagctgg tgtacagcgt ggcaaggggc gactccgcga caaatgaaca ctgggttact aagggcccat gccctgggct ggcgctctga tccctcagca aacgtagatc gtcgagtgcc ccaaaaccca gacgtgagcc cataat.gcca gtcctcaccg aacaaaggcc gaaccacagg ctgacctgcc gggcagccgg ttcctctaca tgctccgtga ccgggtaaat ggctgagctg tggagtctgg ctggattcac tggagtgggt agggccgatt gcctgagagc ttgactactg cggtcttccc gcctggtcaa ccagcggcgt gcgtggtgac acaagcccag caccgtgccc aggaccct acgaagaccc óięćlCclclHCJCC ttgtgcacca tcccagcccc tgtacaccct tggtcaaagg agaacaacta gcaagctcac tgcatgaggc branch ggttttcctc gggaggcgtg cttcagtagt ggcagttata caccatctcc cgaggacacg gggccaggga cctggcgccc ggactacttc gcacaccttc cgtgccctcc caacaccaag agcaccacct catgatctcc cgaggtccag acgggaggag ggactggctg catcgagaaa gcccccatcc cttctacccc caagaccaca cgtggacaag tctgcacaac gttgctcttt gtcgagcctg tatggcatgc tggtatgatg agagacaatt gctgtgtatt accctggtca tgctccagga cccgaaccgg ccagctgtcc agcaacttcg gtggacaaga gtggcaggac cggacccctg ttcaactggt cagttcaaca aacggcaagg accatctcca cgggaggaga agcgacatcg cctcccatgc agcaggtggc cactacacgc taagaggtgt ggaggtccct actgggtccg gaagcaataa ccaagaacac actgtgcgag ccatctcctc gcacctccga tgacggtgtc tacagtcct.c gcacccagac cagttgagcg cgtcagtctt aggtcacgtg acgtggacgg gcacgttccg agtacaagtg aaaccaaagg tgaccaagaa ccgtggagtg tggactccga agcaggggaa agaagagcct ccagtgtcag gagactctcc ccaggctcca acactatgca gctgtatctg agccggactg agcctccacc gagcacagcg gtggaactca aggactctac ctacacctgc caaatgttgt cctcttcccc cgtggtggtg cgtggaggtg tgtggtcagc caaggtctcc gcagccccga ccaggtcagc ggagagcaat cggctccttc cgtcttctca ctccctgtct

120

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240

300

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660

720

780

840

900

960

1020

1080

1140

1.200

1260

1320

1380

1392 &lt;210&gt; 31 &lt;211&gt; 507 &lt;212&gt; DNA &lt;213&gt; Homo sapiens &lt;400&gt; 31 ggcgtggtcc agtagctatg gttatatggt atctccagag gacacggctg tggggccaag agcctgggag gcatgcactg atgatggaag acaattccaa tgtattactg ggaccacggt gtccctgaga ggtccgccag taataaatac gaacacgctg tgcgagaggg caccgtctcc ctctcctgtg gctccaggca tatgcagact tatctgcaaa gcccgtataa tcagcctcca cagcgtctgg aggggctgga ccgtgaaggg tgaacagcct taaccccttg ccaaggaccc attcaccttc gtgggtggca ccgattcacc gagagccgag tatggacgtc atcggtcttc

120

180

240

300

360

PL 214 003 Β1 cccctggcgc aaggactact gtgcacacct cctgctccag tccccgaacc tcccagctgt gagcacctcc ggtgacggtg cctacag gagagcacag tcgtggaact cggccctggg ctgcctcaggctc cagcaggctc gcag

420

480

507 &lt;210 32 &lt;211&gt; 501 &lt;2i2&gt; DNA &lt;213&gt; Homo sapiens &lt;400 32 ggcgtggtcc agtagtcatg gttatatggt atctccagag gacacggctg cagggaaccc gcgccctgct tacttccccg accttcccag agcctgggag gcatccactg atgatggaag acaattccaa tgtattactg tggtcaccgt ccaggagcac aaccggtgac ctgtcctaca gtccctgaga ggtccgccag aaataaagac gaacacgctg tgcgagagtg ctcctcagcc ctccgagagc ggtgtcgtgg g ctctcctgtg gctccaggca tatgcagact tarttgcaaa gccccactgg tccaccaagg acagcggccc aactcaggcg tagcgtctgg aggggctgga ccgtgaaggg tgaacagcct ggccacttga gcccatcggt tgggctgcct ctctgaccag attcatcttc 60 gtgggtggca 120 ccgattcacc 180 gagagccgag 240 ctactggggc 300 cttccccctg 360 ggtcaaggac 420 cggcgtgcac 480

501 &lt;210&gt; 33 &lt;211&gt; 516 &lt;212&gt; DNA &lt;213&gt; Homo sapiens &lt;400 33 tcgggcccag ggctccatca ctggagtgga agtcgagtta gtgactgccg atagacgtct tccgtcttcc tgcctggtca accagcggcg gactggtgaa gcagtggtgg ttgggtacat ccatatcagt cggacacggc ggggccaagg ccctggcgcc aggactactt cgcacacctt gccttcacag tcactactgg ctattacatt agacacgtct cgtgtattat gaccacggtc ctgctccagg ccccgaaccg cccggctgtc atcctgtccc agctggatcc gggaacacct aagaaccagt tgtgcgagag accgtctcct agcacctccg gtgacggtgt ctacaa tcacctgcac gccagcaccc actacaaccc tctccctgaa atagtgggga cagcttccac agagcacagc cgtggaactc tgtctctggt agggaagggc gtccctcaag gctgagctct ctactacggt caagggccca cgccctgggc aggcgccctg

120

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480

516 &lt;210 34 &lt;211&gt; 459 &lt;212&gt; DNA &lt;213&gt; Homo sapiens &lt;400&gt; 34 cctgggaggt atccactggg gatggaagaa aattccaaga tattactgtg gtcaccgtct aggagcacct ccggtgacgg ccctgagact tccgccaggc ataaagacta acacgctgta cgagagtggc cctcagcctc ccgagagcac tgtcgtggaa ctcctgtgca tccaggcaag tgcagactcc tttgcaaatg cccactgggg caccaagggc agcggccctg ctcaggcgct gcgtctggat gggctggagt gtgaagggcc aacagcctga ccacttgact ccatcggtct ggctgcctgg ctgaccagc tcaccttcag gggtggcagt gattcaccat gagccgagga actggggcca tccccctggc tcaaggacta tagtcatggc tatatggtat ctccagagac cacggctgtg gggaaccctg gccctgctcc cttccccgaa

120

180

240

300

360

420

59 &lt;210 35 &lt;211&gt; 503 &lt;212&gt; DNA

GB 214 003 Β1 &lt;213&gt; Homo sapiens agcctgggag gcatgcactg atgatggaag acaattccaa tgtattactg acgtctgggg tcttccccct tggtcaagga gcggcgtgca gtccctgaga ggtccgccag taataaatac gaacacgctg tgcgagagat ccaagggacc ggcgccctgc ctacttcccc cac ctctcctgtg gctccaggca tatgcagact tatctgcaaa ccgaggggag acggtcaccg tccaggagca gaaccggtga cagcgtctgg aggggctgga ccgtgaaggg tgaacagcct ctacccttta tctcctcagc cctccgagag cggtgtcgtg attcaccttc gtgggtggca ccgattcacc gagagccgag ctactactac ctccaccaag cacagcggcc gaactcaggc

120

180

240

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360

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503 &lt;210 36 &lt;211&gt; 451 &lt;212&gt; DNA &lt;213&gt; Homo sapiens &lt;400&gt; 36 ggcgtggtcc agcctgggag gtccctgaga ctctcctgtg cagcgtctgg attcaccttc agr.agctatg gcatgcactg ggtccgccag gctccaggca aggggctgga gtgggtggca gttatatggt atgatggaag tcataaatac tatgcagact ccgtgaaggg ccgattcacc atctccagag acaattccaa gaacacgctg tatctgcaaa tgaacagcct gagagccgag gacacggctg tgtattactg tgcgagaggc gctgtagtag taccagctgc tatggacgtc tggggccaag ggaccacggt caccgtctcc tcagcctcca ccaagggccc atcggtcttc cccctggcgc cctgctccag gagcacctcc gagagcacag cggccctggg ctgcctggtc aaggactact tccccgaacc ggtgacggtg t

120

180

240

300

360

420

451 &lt;210 37 &lt;211&gt; 438 &lt;212&gt; DNA &lt;213&gt; Homo sapiens &lt;22Q&gt;

&lt;221&gt; modifiedjoase &lt;222&gt; (64) &lt;223&gt; a, c, t, g, other or unknown &lt;400 37 gtggtccagc agcngtggca atatggtctg tccagagaca acggctgtgt ggccagggaa ctggcgccct gactacttcc ctgggaggtc tgcactgggt atggaagtca attccaagaa attactgtgc ccctggtcac gctccaggag ccgaaccg cctgagactc ccgccaggct taaatactat cacgctgtat gagaggaact cgtctcctca cacctccgag tcctgtgcag ccaggcaagg gcagactccg ct.gcaaatga atgatagtag gcctccacca agcacagcgg cgtctggatt ggctggagtg tgaagggccg acagcctgag tgggtaccct agggcccatc ccctgggctg caccttcagt ggtggcagtt attcaccatc agccgaggac tgactactgg ggtcttcccc cctggtcaag

120

180

240

300

360

420

439 &lt;210&gt; 38 &lt;211&gt; 562 &lt;212&gt; DNA &lt;213&gt; Homo sapiens &lt;400&gt; 38 tcctgtgcag cgtctggatt caccttcagt tactatggcg tctggggag gcgtggtcca 60

PL 214 003 Β1 gcctgggagg cgtgcactgg tgatggaagt caattccaag gtattattgt ctggggccaa ccccctggcg caaggactac cgtgcacacc tccctgagacccgccagg aataaatact agcacgcaggt gcggcagi ttcccagctg tctcctgtgc ctccaggcaa atgcagactc atctgcaaat cgtattacga tcaccgtctc ggagcacctc cggtgacggt tc agcgtctgga ggggctggag cgtgaagggc gaacagcctg tttttggagt ctcagcctcc cgagagcaca gtcgtggaac ttcaccttca tgggtggcag cgattcacca agagccgagg ggtcggggcg accaagggcc gcggccctgg tcaggcgctc gcagctatgg ttatatggta tctccagaga acacggctgt gtatggacgt catcggtctt gctgcctggt tgaccagcgg

120

ISO

240

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420

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540

562 &lt;210 39 &lt;211&gt; 490 &lt;212&gt; DNA &lt;213&gt; Homo sapiens &lt;400 39 gtccagcctg tatgccatgc tggcatgatg agagacaatt gctgtatatt ggccagggaa ctggcgccct gactacttcc cacaccttcc ggaggtccct actgggtccg gaaataataa ccaagaacac actgtgcgag ccctggtcac gctccaggag ccgaaccggt gagactctcc ccaggctcca atactatgca gctgtatctg agatcagggc cgtctcctca cacctccgag gacggtgtcg tgtgcagcgt ggcaaggggc gagtccgtga caaatgaaca actggctggt gcctccacca agcacagcgg tggaactcag ctggattcac tggagtgggt agggccgatt gcctgagagc acggaggctt agggcccatc ccctgggctg gcgctctgac cttcagt.aac ggtagttatt caccatctcc cgaggacacg tgacttctgg ggtcttcccc cctggtcaag cagcggcgtg

120

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240

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420

480

490 &lt;210&gt;

&lt;211&gt;

&lt;212&gt;

&lt;213&gt;

708

GOUT

Homo sapiens tctcttcctc tccaggcacc gagtattagc cctcatctat gtctgggaca ttactgtcag caaacgaact atctggaact acagtggaag ggacagcaag cgagaaacac aaagagcttc ctgctactct ctgtctttgt agcagcttct ggtgcatcca gacttcactc cagtatggta gtggctgcac gcctctgttg gtggataacg gacagcacct aaagtctacg aacaggggag ggctcccaga ctccagggga tagcctggta gcagggccac tcaccatcag cctcaccctg catctgtctt tgtgcctgct ccctccaatc acagcctcag cctgcgaagt agtgttag

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240

300

360

420

480

540

600

660

708

702 DNA Homo sapiens &lt;400&gt; 41 atggaaaccc cagcgcagct tctcttcctc ctgctactct ggctcccaga taccaccgga60 gaaattgtgt tgacgcagtc tccaggcacc ctgtctttgt ctccagggga aagagccacc120 ctctcctgca ggaccagtgt tagcagcagt tacttagcct ggtaccagca gaaacctggcISO caggctccca ggctcctcat ctatggtgca tccagcaggg ccactggcat cccagacagg240

EN 214 003 Β1 ttcagtggca gattttgcag accaaggtgg gatgagcagt agagaggcca agtgtcacag agcaaagcag agctcgcccg gtgggtctgg tctattactg agatcaagcg tgaaatctgg aagtacagtg agcaggacag actacgagaa tcacaaagag gacagacttc tcagcagtat aactgtggct aactgcctct gaaggtggat caaggacagc acacaaagtc cttcaacagg actctcacca ggcatctcac gcaccatctg gttgtgtgcc aacgccctcc acctacagcc tacgcctgcg ggagagtgtt tcagcagact tcttcatctt tgctgaataa aatcgggtaa tcagcagcac aagtcaccca ag ggagcctgaa cggcggaggg cccgccatct cttctatccc ctcccaggag cctgacgctg tcagggcctg

300

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600

660

702 &lt;210&gt; 42 &lt;211&gt; 417 &lt;212&gt; DNA &lt;213&gt; Homo sapiens &lt;400&gt; 42

<td>ggcaccctgt</td><td>ctttgtctcc</td><td>aggggaaaga</td><td>gccaccctct</td><td>cctgcagggc</td><td>cagtcagagt</td><td> 60</td>

<td>gtcagcagct</td><td>acttagcctg</td><td>gtaccagcag</td><td>aaacctggcc</td><td>aggctcccag</td><td>actcctcatc</td><td> 120</td>

<td>tatggtgcat</td><td>ccagcagggc</td><td>cactggcatc</td><td>ccagacaggt</td><td>tcagtggcag</td><td>tgggtctggg</td><td> 180</td>

<td>acagacttca</td><td>ctctcaccat</td><td>cagcagactg</td><td>gagcctgagg</td><td>attttgcagt</td><td>gtattactgt</td><td> 240</td>

<td>cagcagtatg</td><td>gtaggtcacc</td><td>attcactttc</td><td>ggccctggga</td><td>ccaaagtgga</td><td>tatcaagcga</td><td> 300</td>

<td>actgtggctg</td><td>caccatctgt</td><td>cttcatcttc</td><td>ccgccatctg</td><td>atgagcagtt</td><td>gaaatctgga</td><td> 3 60</td>

<td>actgcctctg</td><td>ttgtgtgcct</td><td>gctgaataac</td><td>ttctatccca</td><td>gagaggccaa</td><td>agtacag</td><td> 417</td>

<td> &lt;210&gt; 43</td><td></td><td></td><td></td><td></td><td></td><td></td>

<td> &lt;211&gt; 705</td><td></td><td></td><td></td><td></td><td></td><td></td>

<td>&lt;212&gt; DNA</td><td></td><td></td><td></td><td></td><td></td><td></td>

<td>&lt;213&gt; Homo</td><td>sapiens</td><td></td><td></td><td></td><td></td><td></td>

<td> &lt;400&gt; 43</td><td></td><td></td><td></td><td></td><td></td><td></td>

<td>atggaaaccc</td><td>cagcgcagct</td><td>tctcttcctc</td><td>ctgctactct</td><td>ggctcccaga</td><td>taccaccgga</td><td> 60</td>

<td>gaaattgtgt</td><td>tgacgcagtc</td><td>tccaggcacc</td><td>ctgtctttgt</td><td>ctccagggga</td><td>aagagccacc</td><td> 120</td>

<td>ctctcctgta</td><td>gggccagtca</td><td>aagtgttagc</td><td>agctacttag</td><td>cctggtacca</td><td>acagaaacct</td><td> 180</td>

<td>ggccaggctc</td><td>ccaggcccct</td><td>catctatggt</td><td>gtatccagca</td><td>gggccactgg</td><td>catcccagac</td><td> 2 4 0</td>

<td>aggttcagtg</td><td>gcagtgggtc</td><td>tgggacagac</td><td>ttcactctca</td><td>ccatcagcag</td><td>actggagcct</td><td> 300</td>

<td>gaagattttg</td><td>cagtgtatta</td><td>ctgtcagcag</td><td>tatggtatct</td><td>caccattcac</td><td>tttcggccct</td><td> 3 60</td>

<td>gggaccaaag</td><td>tggatatcaa</td><td>acgaactgtg</td><td>gctgcaccat</td><td>ctgtcttcat</td><td>cttcccgcca</td><td> 420</td>

<td>tctgatgagc</td><td>agttgaaatc</td><td>tggaactgcc</td><td>tctgttgtgt</td><td>gcctgctgaa</td><td>taacttctat</td><td> 480</td>

<td>cccagagagg</td><td>ccaaagtaca</td><td>gtggaaggtg</td><td>gataacgccc</td><td>tccaatcagg</td><td>taactcccag</td><td> 540</td>

<td>gagagtgtca</td><td>cagagcagga</td><td>cagcaaggac</td><td>agcacctaca</td><td>gcctcagcag</td><td>caccctgacg</td><td> 600</td>

<td>ctgagcaaag</td><td>cagactacga</td><td>gaaacacaaa</td><td>gtctacgcct</td><td>gcgaagtcac</td><td>ccatcagggc</td><td> 660</td>

<td>ctgagctcgc</td><td>ccgtcacaaa</td><td>gagcttcaac</td><td>aggggagagt</td><td>gttag</td><td></td><td> 705</td>

&lt;210&gt; 44 &lt;211&gt; 458 &lt;212&gt; DNA &lt;213&gt; Homo sapiens &lt;400&gt; 44 cagtctccat agtcagagca ttcctgatct ggctctggga tactactgtc atcaaacgaa aaatctggaa cctccctgtc ttaacaccta ctgctacatc caaatttcac aacagagtta ctgtggctgc ctgcctctgt tgcatctgta tttaatttgg cattttgcaa tctcaccatc cagtacccca accatctgtc tgtgtgcctg ggagacagag tatcagcaga agtggggtcc aacagtcttc ttcactttcg ttcatcttcc ctgaataact tcaccatcac aaccagggaa catcaaggtt atcctgaaga gccctgggac cgccatctga tctatcccag ttgccgggca agcccctaac ccgtggcagt ttttgcaact caaagtggat tgagcagttg agaggccaaa

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PL 214 003 Β1 gtacagtgga aggtggataa cgccctccaa tcgggtaa

458 &lt;210&gt; 45 &lt;211&gt; 426 &lt;212&gt; DNA &lt;213&gt; Homo sapiens &lt;400&gt; 45 tctccaggca cagagtatta ctcctcatct gggtctggga tattactgtc atcaagcgaa aaatctggaa gtacag ccctgtcttt gcagcaattt atcgtccatc cagacttcac agcagtatgg ctgtggctgc ctgcctctgt gtctccaggg cttagcctgg cagcagggcc tctcaccatc tacgtcacca accatctgtc tgtgtgcctg gaaagagcca taccagcaga actggcatcc agcagactgg ttcactttcg ttcatcttcc

Ca d L * O dk - * ccctctcctg aacctggcca cagacagttt agcctgagga gccctgggac cgccatctga tctatcccag cagggccagt ggctcccagg cagtggcagt ttttgcatta caaagtgcagat tgagcagcatta caaagtgcaag

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426 &lt;210&gt; 46 &lt;211&gt; 465 &lt;212&gt; DNA &lt;213&gt; Homo sapiens tctccagact cagagcattg ctcatcaagt tctgggacag tactgtcatc aaacgaactg tctggaactg cagtggaagg ttcagtctgt gtagtagctt atgcttccca atttcaccct agagtagtag tggctgcacc cctctgttgt tggataacgc gactccaaag acattggtat gtccttctct caccatcaat tttaccgctc atctgtcttc gtgcctgctg cctccaatcg gagaaagtca cagcagaaac ggggtcccct agcctggaag actttcggcg atcttcccgc aataacttct ggtaactccc ccatcacctg cagatcagtc cgaggttcag ctgaagatgc gagggaccaa catctgatga atcccaaaga aggag ccgggccagt tccaaagctc tggcagtgga tgcaacgtat ggtggagatc gcagttgaaa ggccaaagta

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465 &lt;210&gt; 47 &lt;211&gt; 440 &lt;212&gt; DNA &lt;213&gt; Homo sapiens &lt;400&gt; 47 cagtctccag agtcagagtg ctcctcatct gggtctggga tattactgtc atcaagcgaa aaatctggaa gtacagtgga gcaccctgtc tcagcagcta atggtgcatc cagacttcac aacagtatgg ctgtggctgc ctgcctctgt aaggtggata tttgtctcca cttagcctgg cagcagggcc tctcaccatc taggtcacca accatctgtc tgtgtgcctg ggggaaaga.g taccagcaga actggcatcc agcagactgg ttcactttcg ttcatcttcc ctgaataact ccaccctctG aacctggcca cagacaggtt agcctgagga gccctgggac cgccatctga tctatcccag ctgcagggcc ggctcccagg cagtggcagt ttttgcagtg caaagtagat tgagcagttg agaggccaaa

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440 &lt;210&gt; 48 &lt;211&gt; 417 &lt;212&gt; DNA &lt;213&gt; Homo sapiens &lt;400&gt; 48 ccatcctccc tgtctgcatc tgtaggagac agagtcacca tcacttgccg ggcaagtcag 60

EN 214 003 Β1 agcattaaca atctatgctg gggacagatt tgtcaac.agt cgaactgtgg ggaactgcct gctatttaga catccagttt tcactctcac attacagtac ctgcaccatc ctgttgtgtg ttggtatcag gcaaagtggg catcagcagt tccattcact tgtcttcatc cctgctgaat cagaaaccag gtcccatcaa ct.gcaacctg ttcggccctg ttcccgccat aacttctatc ggaaagcccc ggttcagtgg aagattttgc ggaccaaagt ctgatgagca caaactcctg cagtggatct aacttactac ggaaatcaaa gttgaaatct

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360 ccagagaggc caaagta

417 &lt;210&gt; 49 &lt;211&gt; 402 &lt;212&gt; DNA &lt;213&gt; Homo sapiens &lt;220&gt;

&lt;221&gt; modified base &lt;222&gt; (207) &lt;223&gt; a, c, t, g, other or unknown &lt;400&gt; 49 acccagtctc gcaagtcaga aagttcctga agtggatctg acttactact gatatcaaac ttgaaatctg catcctccct acattagcag tctatgttgc ggccagattt gtcaacagag gaactgtggc gaactgcctc gtctgcatct gtatttaaat atctattttg cactctnacc ttacagtacc tgcaccatct tgttgtgtgc gtaggagaca tggtatcaac caaagtgggg atcagcagtc ccattcactt gtcttcatct ctgctgaata gagtcaccat agaaaccagg tcccatcagg tgcaacctga tcggccctgg tcccgccatc ac cacttgccgg gaaagcccct gttcagtgcc agattttgca gaccaaagtg tgatgagcag

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<td> &lt;210&gt; &lt;211&gt; &lt;212&gt; &lt;213&gt;</td><td>50 451 GOUT Homo sapiens</td>

<td> &lt;400&gt;</td><td> 50</td>

acccagtctc gcaagtcaga agggtcctga agtggatctg acttactact gatatcgaac ttgaaatctg aaagtacagt catcctccct gcatttgcaa tctatgctgc ggacagattg gtcaacagag gaactgtggc gaactgcctc ggaaggtgga gtctgcatct ctatttaaat atccagtttg cactctcacc ttacactacc tgcaccatct tgttgtgtgc taacgcctat gtaggagaca tggtatcagc caaggtgggg atcagcagtc ccattcactt gtcttcatct ctgctgaata gagtcaccat agaaaccagg tcccgtcaag tgcaacctga tcggccctgg tcccgccatc acttctatcc cacttgccgg aaaagcccct gttcagtggc agattttgca gaccagagtg tgatgagcag cagagaggcc

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451 &lt;210&gt; 51 &lt;211&gt; 419 &lt;212&gt; DNA &lt;213&gt; Homo sapiens &lt;400&gt; 51 ccactctccc tgcccgtcac ccttggacag ccggcctcca tctcctgcag gtctagtcaa60 agcctcgtat acagtgatgg aaacacctac ttgaattggt ttcagcagag gccaggccaa120 tctccaaggc gcctaattta taaggtttct aactgggact ctggggtccc agacagattc180 agcggcagtg ggtcaggcac tgatttcaca ctgaaaatca gcagggtgga ggctgaggat240 gttggggttt attactgcat gcaaggttca. cactggcctc cgacgttcgg ccaagggacc300 aaggtggaaa tcaaacgaac tgtggctgca ccatctgtct tcatcttccc gccatctgat360 gagcagttga aatctggaac tgcctctgtt gtgtgcc.tgc tgaatacactact4

GB 214 003 Β1 &lt;210&gt; 52 &lt;211&gt; 419 &lt;212&gt; DNA &lt;213&gt; Homo sapiens &lt;400&gt; 52 cctggagagc cggcttccat ctcttgcagg tctagtcaga gcctcctgca tagtaatgga60 tacaactatt tggattggta cctgcagaag ccaggacagt ctccacagct cctgatctat120 ttgggttcta atcgggcctc cggggtccct gacaggttca gtggcagtgg atcaggcaca180 gattttacao tgaaactcag cagagtggag gctgaggatg ttggggttta ttactgcatg240 caagctctac aaactcctct cactttcggc ggagggacca aggtggagat caaacgaact300 gtggctgcac catctgtctt catcttcccg ccatctgatg agcagttgaa atctggaact360 gcctctgttg tgtgcctgct gaataacttc tatcccagar aggccaaagt acattccat419 &lt;210&gt; 53 &lt;2LL&gt; 1392 &lt;212&gt; DNA &lt;213&gt; Homo sapiens &lt;400&gt; 53 atggagtttg gtgcagctgg tgtgtagcgt ggcaaggggc gactccgtga caaatgaaca ttcggtcctt aagggcccat gccctgggct ggcgctctga tccctcagca aacgtagatc gtcgagLgcc ccaaaaccca gacgtgagcc cataatgcca gtcctcaccg aacaaaggcc gaaccacagg ctgacctgcc gggcagccgg ttcctctaca tgctccgtga ccgggtaaat ggctgagctg tggagtctgg ctggattcac tggagtgggt agggccgatt gcctgagagc ttgactactg cggtcttccc gcctggtcaa ccagcggcgt gcgtggtgac acaagcccag caccgtgccc aggacaccct acgaagaccc agacaaagcc ttgtgcacca tcccagcccc tgtacaccct tggtcaaagg agaacaacta gcaagctcac tgcatgaggc branch ggttttcctc gggaggcgtg cttcagtagc ggcagttata caccatctcc cgaggacacg gggccaggga cctggcgccc ggactacttc gcacaccttc cgtgccctcc caacaccaag agcaccacct catgatctcc cgaggtccag acgggaggag ggactggctg catcgagaaa gcccccatcc cttctacccc caagaccaca cgtggacaag tctgcacaac gttgctcttt gtccagcctg catggcatgc tggtatgatg agagacaatt gctgtgtatt accctggtca tgctccagga cccgaaccgg ccagctgtcc agcaacttcg gtggacaaga gtggcaggac cggacccctg ttcaactggt cagttcaaca aacggcaagg accatctcca cgggaggaga agcgacatcg cctcccatgc agcaggtggc cactacacgc taagaggtgt ggaggtccct actgggtccg gaagaaataa ccaagaacac actgtgcgag ccgtctcctc gcacctccga tgacggtgtc tacagtcctc gcacccagac cagttgagcg cgtcagtctt aggtcacgtg acgtggacgg gcacgttccg agtacaagtg aaaccaaagg tgaccaagaa ccgtggagtg tggactccga agcaggggaa agaagagcct ccagtgtcag gagactctcc cęaggctcca atactatgca gctgtttctg aggaggtcac agcctccacc gagcacagcg gtggaactca aggactctac ctacacctgc caaatgttgt cctcttcccc cgtggtggtg cgtggaggtg tgtggtcagc caaggtctcc gcagccccga ccaggtcagc ggagagcaat cggctccttc cgtcttctca ctcc

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1392 &lt;210&gt; 54 &lt;211&gt; 1999 &lt;212&gt; DNA &lt;213&gt; Homo sapiens &lt;400&gt; 54 atggagtttg ggctgagctg ggttttcctc gttgctcttt taagaggtgt ccagtgtcag60 gtgcagctgg tggagtctgg gggaggcgtg gtccagcctg ggaggtccct gagactctcc120 tgtgtagcgt ctggattcac cttcagtagc catggcatgc actgggtccg ccaggctcca180 ggcaaggggc tggagtgggt ggcagttata tggtatgatg gaagaaataa atactatgca240

EN 214 003 Β1 gactccgtga caaatgaaca ttcggtcctt aagggcccat gccctgggct ggcgctctga tcccLcagca aacgtagatc cagggaggga ctgtgcagcc tctgcccgcc gcaggcacag tcagacctgc ggccaaactg ccaatcttct cagcccaggc agggacaggc gtggcaggac cggacccctg ttcaactggt cagttcaaca aacggcaagg accatctcca gctcggccca gccccgagaa ggtcagcctg gagcaatggg ctccttcttc cttctcatgc cctgtctccg agggccgatt gcctgagagc ttgactactg cggtcttccc gcctggtcaa ccagcggcgt gcgtggtgac acaagcccag gggtgtctgc ccagcccagg ccactcatgc gctgggtgcc caaaagccat tccactccct ctctgcagag ctcgccctcc cccagctggg cgtcagtctt aggtcacgtg acgtggacgg gcacgttccg agtacaagtg aaaccaaagg ccctctgccc ccacaggtgt acctgcctgg cagccggaga ctctacagca tccgtgatgc ggtaaatga caccatctcc cgaggacacg gggccaggga cctggcgccc ggactacttc gcacaccttc cgtgccctcc caacaccaag tggaagccag gcagcaaggc tcagggagag cctaccccag atccgggagg cagctcggac cgcaaatgtt agctcaaggc tgctgacacg cctcttcccc cgtggtggtg cgtggaggtg tgtggtcagc caaggtctcc tgggacccgc tgggagtgac acaccctgcc tcaaaggctt acaactacaa agctcaccgt atgaggctct agagacaatt gctgtgtatt accctggtca tgctccagga cccgaaccgg ccagctgtcc agcaacttcg gtggacaaga gctcagccct aggccccatc ggtcttctgg gcccttcaca accctgcccc accttctctc gtgtcgagtg gggacaggtg tccacctcca ccaaaaccca gacgtgagcc C3.tZ3ćLtlCJCC3 gtcctcaccg aacaaaggcc ggggtatgag cgctgtgcca cccatcccgg ctaccccagc gaccacacct ggacaagagc gcacaaccac ccaagaacac actgtgcgag ccgtctcctc gcacctccga tgacggtgtc tacagtcctc gcacccagac cagttggtga cctgcctgga tgtctcctca ctttttccac cacaggggca tgacctaagc ctcccagatc cccaccgtgc ccctagagta tctcttcctc aggacaccct acgaagaccc agacaaagcc ttgtgcacca tcccagcccc ggccacatgg acctctgtcc gaggagatga gacatcgccg cccatgctgg aggtggcagc tacacgcaga gctgtttctg aggaggtcac agctagcacc gagcacagcg gtggaactca aggactctac ctacacctgc gaggccagct cgcaccccgg cccggaggcc caggctccag ggtgcttggc cgaccccaaa cąagtaactc ccaggtaagc gcctgcatcc agcaccacct catgatctcc cgaggtccag acgggaggag ggactggctg catcgagaaa acagaggccg ctacagggca ccaagaacca tggagtggga actccgacgg aggggaacgt agagcctctc

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<td> &lt;210&gt;</td><td> 55</td>

<td> &lt;211&gt;</td><td> 1392</td>

<td> &lt;212&gt;</td><td>GOUT</td>

<td> &lt;213&gt;</td><td>Homo</td>

<td> &lt;400&gt;</td><td> 55</td>

sapiens atggagtttg gtgcagctgg tgtgtagcgt ggcaaggggc gactccgtga caaatgaaca ttcggtcctt aagggcccat gccctgggct ggcgctctga tccctcagca aacgtagatc gtcgagtgcc ccaaaaccca gacgtgagcc cataatgcca gtcctcaccg aacaaaggcc gaaccacagg ctgacctgcc ggctgagctg tggagtctgg ctggattcac tggagtgggt agggccgatt gcctgagagc ttgactactg cggtcttccc gcctggtcaa ccagcggcgt gcgtggtgac acaagcccag caccgtgccc aggacaccct acgaagaccc agacaaagcc ttgtgcacca tcccagcccc tgtacaccct tggtcaaagg ggttttcctc gggaggcgtg cttcagtagc ggcagttata caccatctcc cgaggacacg gggccaggga cctggcgccc ggactacttc gcacaccttc cgtgccctcc csscsccssę agcaccacct catgatctcc cgaggtccag acgggaggag ggactggctg catcgagaaa gcccccatcc cttctacccc gttgctcttt gtccagcctg catggcatgc tggtatgatg agagacaatt gctgtgtatt accctggtca tgctccagga cccgaaccgg ccagctgtcc agcaacttcg gtggacaaga gtggcaggac cggacccctg ttcaactggt cagttccaaa aacggcaagg accatctcca cgggaggaga agcgacatcg taagaggtgt ggaggtccct actgggtccg gaagaaataa ccaagaacac actgtgcgag ccgtctcctc gcacctccga tgacggtgtc tacagtcctc gcacccagac cagttgagcg cgtcagtctt aggtcacgtg acgtggacgg gcacgttccg agtacaagtg aaaccaaagg tgaccaagaa ccgtggagtg ccagtgtcag gagactctcc ccaggctcca atactatgca gctgtttctg aggaggtcac agcctccacc gagcacagcg gtggaactca aggactctac ctacacctgc caaatgttgt cctcttcccc cgtggtggtg cgtggaggtg tgtggtcagc caaggtctcc gcagccccga ccaggtaagc ggagagcaat

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PL 214 003 Β1 gggcagccgg ttcctctaca tgctccgtga ccgggtaaat.

&lt;210&gt; 56 &lt;211&gt; 708 &lt;212&gt; DNA &lt;213&gt; Homo &lt;400&gt; 56 atggaaaccc gaaattgtgt ctctcctgca cctggccagg gacaggttca cctgaagatt caagggacca ccatctgatg tatcccagag caggagagtg acgctgagca ggcctgagct aęaacaacta gcaagctcac tgcatgaggc branch sapiens cagcgcagct tgacgcagtc gggccagtca ctcccaggct gtggcagtgg ttgcagtgta aggtggaaat agcagttgaa aggccaaagt tcacagagca aagcagacta cgcccgtcac caagaccaca cgtggacaag tctgcacaac tctcttcctc tccaggcacc gagtattagc cctcatctat gtctgggaca ttactgtcag caaacgaact atctggaact acagtggaag ggacagcaag cgagaaacac aaagagcttc cctcccatgc agcaggtggc cactacacgc tggactccga agaagagcct cggctccttc cgtcttctca ctccctgtct

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1392 ctgctactct ctgtctttgt agcagcttct ggtgcatcca gacttcactc cagtatggta gtggctgcac gcctctgttg gtggataacg gacagcacct aaagtctacg aacaggggag ggctcccaga ctccagggga tagcctggta gcagggccac tcaccatcag cctcaccctg catctgtctt tgtgcctgct ccctccaatc acagcctcag cctgcgaagt agtgttag taccaccgga SćlCJełęCCclCC ccagcagaga tggcatccca cagactggag gacgttcggc catcttcccg gaataacttc gggtaactcc cagcaccctg cacccatcag

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600

660

708 &lt;210&gt; 57 &lt;211&gt; 1395 &lt;212&gt; DNA &lt;213&gt; Homo &lt;400&gt; 57 atggagtttg gtgcagctgg tgtacagcgt ggcaaggggc gactccgtga caaatgaaca ctggggtcct accaagggcc gcggccctgg tcaggcgctc tactccctca tgcaacgtag tgtgtcgagt cccccaaaac gtggacgtga gtgcataatg agcgtcctca tccaacaaag cgagaaccac agcctgacct aatgggcagc ttcttcctct tcatgctccg tctccgggta sapiens ggctgagctg ggttttcctc gttgctcttt taagaggtgt ccagtgtcag 60 tggagtctgg gggaggcgtg gtccagcctg ggaggtccct gagactctcc 120 ctggattcac cttcagtaac tatggcatgc actgggtccg ccaggctcca 180 tggagtgggt ggcagttata tggtatgatg gaagtaataa acactatgga 240 agggccgatt caccatctcc agtgacaatt ccaagaacac gctgtatctg 300 gcctgagagc cgaggacacg gctgtgtatt actgtgcgag aggagagaga 360 actttgacta ctggggccag ggaaccctgg tcaccgtctc ctcagcctcc 420 catcggtctt ccccctggcg ccctgctcca ggagcacctc cgagagcaca 480 gctgcctggt caaggactac ttccccgaac cggtgacggt gtcgtggaac 540 tgaccagcgg cgtgcaca cc ttcccagctg tcctacagtc ctcaggactc 6C0 gcagcgtggt gaccgtgccc tccagcaact tcggcaccca gacctacacc 660 atcacaagcc cagcaacacc aaggtggaca agacagttga gcgcaaatgt 720 gcccaccgtg cccagcacca cctgtggcag gaccgtcagt cttcctcttc 780 ccaaggacac cctcatgatc tcccggaccc ctgaggtcac gtgcgtggtg 840 gccacgaaga ccccgaggtc cagttcaact ggtacgtgga cggcgtggag 900 ccaagacaaa gccacgggag gagcagttca acagcacgtt ccgtgtggtc 960 ccgttgtgca ccaggactgg ct.gaacggca aggagtacaa gtgcaaggtc 1020 gcctcccagc ccccatcgag aaaaccatct ccaaaaccaa agggcagccc 1080 aggtgtacac cctgccccca tcccgggagg agatgaccaa gaaccaggtc 1140 gcctggtcaa aggcttctac cccagcgaca tcgccgtgga gtgggagagc 1200 cggagaacaa ctacaagacc acacctccca tgctggactc cgacggctcc 1260 acagcaagct caccgtggac aagagcaggt ggcagcaggg gaacgtcttc 1320 tgatgcatga ggctctgcac aacagcagcac aacagcag1380 cggactgcac aacagacagactaca 1320

GB 214 003 Β1 &lt;210&gt; 58 &lt;211&gt; 702 &lt;212&gt; DNA &lt;213&gt; Homo sapiens &lt;400&gt; 58 atggaaaccc gaaartgtgt ctctcctgca caggctccca ttcagtggca gattttgcag accaaggtgg gatgagcagt agagaggcca agtgtcacag agcaaagcag agctcgcccg cagcgcagct tgacgcagtc ggaccagtgt ggctcctcat gtgggtctgg tctattactg agatcaagcg tgaaatctgg aagtacagtg agcaggacag actacgagaa tcacaaagag tctcttcctc tccaggcacc tagcagcagt ctatggtgca gacagacttc tcagcagtat aactgtggct aactgcctct gaaggtggat caaggacagc acacaaagtc cttcaacagg ctgctactct ctgtctttgt tacttagcct tccagcaggg actctcacca ggcatctcac gcaccatctg gttgtgtgcc aacgccctcc acctacagcc tacgcctgcg ggagagtgtt ggctcccaga ctccagggga ggtaccagca ccactggcat tcagcagact ccttcacttt tcttcatctt tgctgaataa aatcgggtaa tcagcagcac aagt caccca ag taccaccgga aagagccacc gaaacctggc cccagacagg ggagcctgaa cggcggaggg cccgccatct cttctatccc ctcccaggag cctgacgctg tcagggcctg

120

190

240

300

360

420

480

540

600

660

702 &lt;210&gt; 59 &lt;211&gt; 1392 &lt;212&gt; DNA &lt;213&gt; Homo sapiens &lt;400&gt; 59 atggagtttg gtgcagctgg tgtacagcgt ggcaaggggc gactccgcga caaatgaaca ctgggttact aagtaggcccat gccctggctca a gccctggctca gccgcggctca acgtaggctccc gccctggctca gccgcggctca

C · ™. Those dd 6- C5 · C &lt;· gacgtgagcc cataatgcca gtcctcaccg aacaaaggcc gaaccacagg ctgacctgcc gggcagccgg ttcctctaca tgctccgtga ccgggtaaat ggctgagctg tggagtctgg ctggattcac tggagtgggt agggccgatt gcctgagagc ttgactactg cggtcttccc gcctggtcaa ccagcggcgt gcgtggtgac acaagcccag caccgtgccc aggacaccct acgaagaccc agacaaagcc ttgtgcacca tcccagcccc tgtacaccct tggtcaaagg agaacaacta gcaagctcac tgcatgaggc branch ggttttcctc gggaggcgtg cttcagtagt ggcagttata caccatctcc cgaggacacg gggccaggga cctggcgccc ggactacttc gcacaccttc cgtgccctcc caacaccaag agcaccacct catgatctcc cgaggtccag acgggaggag ggactggctg catcgagaaa gcccccatcc cttctacccc caagaccaca cgtggacaag tctgcacaac gttgctcttt gtcgagcctg tatggcatgc tggtatgatg agagacaatt gctgtgtatt accctggtca tgctccagga cccgaaccgg ccagctgtcc agcaacttcg gtggacaaga gtggcaggac cggacccctg ttcaactggt cagttcaaca aacggcaagg accatctcca cgggaggaga agcgacatcg cctcccatgc agcaggtggc cactacacgc taagaggtgt ggaggtccct actgggtccg gaagcaataa ccaagaacac actgtgcgag ccgtctcctc gcacctccga tgacggtgtc tacagtcctc gcacccagac cagttgagcg cgtcagtctt aggtcacgtg acgtggacgg gcacgttccg agtacaagtg aaaccaaagg tgaccaagaa ccgtggagtg tggactccga agcaggggaa agaagagcct ccagtgtcag gagactctcc ccaggctcca acactatgca gctgtatotg agccggactg agcctccacc gagcacagcg gtggaactca aggactctac ctacacctgc caaatgttgt cctcttcccc cgtggtggtg cgtggaggtg tgtggtcagc caaggtctcc gcagccccga ccaggtcagc ggagagcaat cggctccttc cgtcttctca ctccctgtct

120

180

240

300

60

420

480

540

600

660

720

780

840

900

960

1020

1080

1140

1200

1260

1320

1380

1392

<td> &lt;210&gt;</td><td> 60</td>

<td> &lt;211&gt;</td><td> 705</td>

<td> &lt;212&gt;</td><td>GOUT</td>

<td> &lt;213&gt;</td><td>Homo</td>

sapiens &lt;400&gt; 60

GB 214 003 Β1 atggaaaccc gaaattgtgt ctctcctgta ggccaggctc aggttcagtg gaagattttg gggaccaaag tctgatgagc cccagagagg gagagtgtca ctgagcaaag ctgagctcgc cagcgcagct tgacgcagtc gggccagtca ccaggcccct gcagtgggtc cagtgtatta tggatatcaa agttgaaatc ccaaagtaca cagagcagga cagactacga ccgtcacaaa tctcttcctc tccaggcacc aagtgttagc catctatggt tgggacagac ctgtcagcag acgaactgtg tggaactgcc gtggaaggtg cagcaaggac gaaacacaaa gagcttcaac ctgctactct ctgt No, 1t. agctacttag gt gtatccagca ttcactctca tatggtatct gctgcaccat tctgttgtgt gataacgccc agcacctaca gtctacgcct aggggagagt ggctcccaga ctccagggga cctggtacca gggccactgg ccatcagcag caccattcac ctgtcttcat gcctgctgaa tccaatcggg gcctcagcag gcgaagtcac gttag taccaccgga aagagccacc acagaaacct catcccagac actggagcct tttcggccct cttcccgcca taacttctat taactcccag caccctgacg Gcatcagggc

120

180

240

300

360

420

480

540

600

660

705 &lt;210&gt; 61 &lt;211&gt; 1413 &lt;212&gt; DNA &lt;213&gt; Homo sapiens &lt;400&gt; 61 atggagtttg gtgcagctgg tgtgcagcgt ggcaaggggc gac.tccgtga caaatgaaca ggagctaccc accgtctcct agcacctccg gtgacggtgt ctacagtcct ggcacccaga acagttgagc ccgtcagtct gagqtcacgt tacgtggacg agcacgctcc gagtacaagt aaaaccaaag atgaccaaga gccgtggagt ctggactccg cageagggga cagaagagcc ggctgagctg tggagtctgg ctggattcac tggagtgggt agggccgatt gcctgagagc tttactacta cagcctccac agagcacagc cgtggaactc caggactcta cctacacctg gc.aaatgttg tcctcttccc gcgtggtggt gcgtggaggt gtgtggtcag gcaaggtctc ggcagccccg accaggtcag gggagagcaa acggctcctt acgtcttctc tctccctgtc ggttttcctc gggaggcgtg cttcagtagc ggcagttata caccatctcc cgaggacacg ctactacggt caagggccca ggccctgggc aggcgctctg ctccctcagc caacgtagat tgtcgagtgc cccaaaaccc ggacgtgagc gcataatgcc cgtcctcacc oaacaaaggc agaaccacag cctgacctgc tgggcagccg cttcctctac atgctccgtg tccgggtaaa gttgctcttt gtccagcctg tatggcatgc tggtatgatg agagacaatt gctgtgtatt atggacgtct tcggtcttcc tgcctggtca accagcggcg agcgtggtga cacaagccca ccaccgtgcc aaggacaccc cacgaagaGC aagacaaagc gttgtgcacc ctcccagccc gtgtacaccc crggtcaaag gagaacaact agcaagctca atgcatgagg tga taagaggtgt ggaggtccct actgggtccg gaagtaataa ccaagaacac actgtgcgag ggggccaagg ccctggcgcc aggactactt tgcacacctt ccgtgccctc gcaacaccaa cagcaccacc tcatgatctc CGgaggtcca cacgggagga aggactggct ccatcgagaa tgcccccatc gcttctaccc acaagaccac ccgtggacaa ctctgcacaa ccagtgtcag gagactctcc ccaggctcca atactatgca gctgtatcLg agatccgagg gaccacggtc ctgctccagg ccccgaaccg cccagctgtc cagcaacttc ggtggacaag tgtggcagga ccggacccct gttcaactgg gcagttcaac gaacggcaag aaccatctcc ccgggaggag cagcgacatc acctcccatg gagcaggtgg ccactacacg

120

180

240

300

360

420

480 540

600 660

720

780 840

900 960 1020 1080

114 0 1200 1260 1320 1380 1413 &lt;210&gt; 62 &lt;211&gt; 714 &lt;212&gt; DNA &lt;213&gt; Homo sapiens &lt;400&gt; 62 atggacatga agatgtgaca gtcaccatca ccatcaaggt caacctgaag gggtccccgc tccagatgac cttgccgggc aagcccctaa tcagtggcag atiUgcaac tcagctcctg ccagtctcca aagtcagagc actcctgatc tggatctggg t tactactgt gggctcctgc tcctccctgt attaacagct tatgctgcat acagatttca caacagtatt tactctggct ctgcatctgt atttagattg ccagtttgca ctctcaccat acagtactcc ccgaggtgcc aggagacaga gtatcagcag aagtggggtc cagcagtctg attcactttc

120

180

240

300

360

GB 214 003 Β1 ggccctggga ccgccatctg ttctatccca tcccaggaga ctgacgctga cagggcctga ccaaagtgga atgagcagtt gagaggccaa gtgtcacaga gcaaagcaga gctcgcccgt aatcaaacga gaaatctgga agtacagtgg gcaggacagc ctacgagaaa cacaaagagc actgtggctg actgcctctg aaggtggata aaggacagca cacaaagtct ttcaacaggg caccatctgt ttgtgtgcct acgccctcca cctacagcct acgcctgcga gagagtgtta cttcatcttc 420 gctgaataac 480 atcgggtaac 540 cagcagcacc 600 agtcacccat 660 gtga 714 &lt;210&gt; 63 &lt;211&gt; 463 &lt;212&gt; PRT &lt;213&gt; Homo sapiens &lt;400&gt; 63

<td>Underworld 1</td><td>Glu</td><td>Phe</td><td colspan="2">Gly Leu 5</td><td>Cheese</td><td colspan="2">Trp val</td><td>Phe</td><td>Leu 10</td><td colspan="2">Val Ala</td><td>Leu</td><td>Leu</td><td>Arg 15</td><td>Gly</td>

<td>Val</td><td>Gin</td><td>Cys</td><td>Gin</td><td>Val</td><td>Gin</td><td>Leu</td><td>Val</td><td>Glu</td><td>Cheese</td><td>dy</td><td>Gly</td><td>dy</td><td>Val</td><td>Val</td><td>Gin</td>

<td></td><td></td><td></td><td> 20</td><td></td><td></td><td></td><td></td><td> 25</td><td></td><td></td><td></td><td></td><td> 30</td><td></td><td></td>

<td>Pro</td><td>Gly</td><td>Arg</td><td>Cheese</td><td>Leu</td><td>Arg</td><td>Leu</td><td>Cheese</td><td>Cys</td><td>Val</td><td>Ala</td><td>Cheese</td><td>Gly</td><td>Phe</td><td>Thr</td><td>Phe</td>

<td></td><td></td><td> 35</td><td></td><td></td><td></td><td></td><td> 40</td><td></td><td></td><td></td><td></td><td> 45</td><td></td><td></td><td></td>

<td>Cheese</td><td>Cheese</td><td>His</td><td>Gly</td><td>Underworld</td><td>His</td><td>Trp</td><td>Val</td><td>Arg</td><td>Gin</td><td>Ala</td><td>Pro</td><td>Gly</td><td>Lys</td><td>Gly</td><td>Leu</td>

<td></td><td> 50</td><td></td><td></td><td></td><td></td><td> 55</td><td></td><td></td><td></td><td></td><td> 60</td><td></td><td></td><td></td><td></td>

<td>Glu</td><td>Trp</td><td>Val</td><td>Ala</td><td>Val</td><td>How much</td><td>Trp</td><td>Tyr</td><td>Asp</td><td>Gly</td><td>Arg</td><td>Asn</td><td>Lys</td><td>Tyr</td><td>Tyr</td><td>Ala</td>

<td> 65</td><td></td><td></td><td></td><td></td><td> 70</td><td></td><td></td><td></td><td></td><td> 7 5</td><td></td><td></td><td></td><td></td><td> 80</td>

<td>ASp</td><td>Cheese</td><td>Val</td><td>Lys</td><td>Gly</td><td>Arg</td><td>Phe</td><td>Thr</td><td>How much</td><td>Cheese</td><td>Arg</td><td>Asp</td><td>Asn</td><td>Cheese</td><td>Lys</td><td>Asn</td>

<td></td><td></td><td></td><td></td><td> 85</td><td></td><td></td><td></td><td></td><td> 90</td><td></td><td></td><td></td><td></td><td> 95</td><td></td>

<td>Thr</td><td>Leu</td><td>Phe</td><td>Leu</td><td>Gin</td><td>Underworld</td><td>Asn</td><td>Cheese</td><td>Leu</td><td>Arg</td><td>Ala</td><td>Glu</td><td>Asp</td><td>Thr</td><td>Ala</td><td>Val</td>

<td></td><td></td><td></td><td> 100</td><td></td><td></td><td></td><td></td><td> 105</td><td></td><td></td><td></td><td></td><td> 110</td><td></td><td></td>

<td>Tyr</td><td>Tyr</td><td>Cys</td><td>Ala</td><td>Arg</td><td>Gly</td><td>Gly</td><td>His</td><td>Phe</td><td>Gly</td><td>Pro</td><td>Phe</td><td>Asp</td><td>Tyr</td><td>Trp</td><td>Gly</td>

<td></td><td></td><td> 115</td><td></td><td></td><td></td><td></td><td> 120</td><td></td><td></td><td></td><td></td><td> 12 5</td><td></td><td></td><td></td>

<td>Gin</td><td>Gly</td><td>Thr</td><td>Leu</td><td>Val</td><td>Thr</td><td>Val</td><td>Cheese</td><td>Cheese</td><td>Ala</td><td>Cheese</td><td>Thr</td><td>Lys</td><td>Gly</td><td>Pro</td><td>Cheese</td>

<td></td><td> 130</td><td></td><td></td><td></td><td></td><td> 135</td><td></td><td></td><td></td><td></td><td> 140</td><td></td><td></td><td></td><td></td>

<td>Val</td><td>Phe</td><td>Pro</td><td>Leu</td><td>Ala</td><td>Pro</td><td>Cys</td><td>Cheese</td><td>Arg</td><td>Cheese</td><td>Thr</td><td>Cheese</td><td>Glu</td><td>Cheese</td><td>Thr</td><td>Ala</td>

<td> 145</td><td></td><td></td><td></td><td></td><td> 150</td><td></td><td></td><td></td><td></td><td> 155</td><td></td><td></td><td></td><td></td><td> 160</td>

<td>Ala</td><td>Leu</td><td>Gly</td><td>Cys</td><td>Leu</td><td>Val</td><td>Lys</td><td>Asp</td><td>Tyr</td><td>Phe</td><td>Pro</td><td>Glu</td><td>Pro</td><td>Val</td><td>Thr</td><td>val</td>

<td></td><td></td><td></td><td></td><td> 1 65</td><td></td><td></td><td></td><td></td><td> 170</td><td></td><td></td><td></td><td></td><td> 175</td><td></td>

<td>Cheese</td><td>Trp</td><td>Asn</td><td>Cheese</td><td>Gly</td><td>Ala</td><td>Leu</td><td>Thr</td><td>Cheese</td><td>Gly</td><td>Val</td><td>His</td><td>Thr</td><td>Phe</td><td>Pro</td><td>Ala</td>

<td></td><td></td><td></td><td> 180</td><td></td><td></td><td></td><td></td><td> 185</td><td></td><td></td><td></td><td></td><td> 190</td><td></td><td></td>

<td>Val</td><td>Leu</td><td>Gin</td><td>Cheese</td><td>Cheese</td><td>Gly</td><td>Leu</td><td>Tyr</td><td>Cheese</td><td>Leu</td><td>Cheese</td><td>Cheese</td><td>Val</td><td>val</td><td>Thr</td><td>Val</td>

<td></td><td></td><td> 195</td><td></td><td></td><td></td><td></td><td> 200</td><td></td><td></td><td></td><td></td><td> 205</td><td></td><td></td><td></td>

<td>Pro</td><td>Cheese</td><td>Cheese</td><td>Asn</td><td>Phe</td><td>Gly</td><td>Thr</td><td>Gin</td><td>Thr</td><td>Tyr</td><td>Thr</td><td>Cys</td><td>Asn</td><td>val</td><td>Asp</td><td>His</td>

<td></td><td> 210</td><td></td><td></td><td></td><td></td><td> 215</td><td></td><td></td><td></td><td></td><td> 220</td><td></td><td></td><td></td><td></td>

<td>Lys</td><td>Pro</td><td>Cheese</td><td>Asn</td><td>Thr</td><td>Lys</td><td>Val</td><td>Asp</td><td>Lys</td><td>Thr</td><td>Val</td><td>Glu</td><td>Arg</td><td>Lys</td><td>Cys</td><td>Cys</td>

<td> 225</td><td></td><td></td><td></td><td></td><td> 230</td><td></td><td></td><td></td><td></td><td> 235</td><td></td><td></td><td></td><td></td><td> 240</td>

PL 214 003 Β1

<td>Va.l</td><td>Glu</td><td>Cys</td><td>Pro</td><td>Pro 245</td><td>Cys</td><td>Pro</td><td>Ala</td><td>Pro</td><td>Pro 250</td><td>Val</td><td>Ala</td><td>Gly</td><td>Pro</td><td>Cheese 255</td><td>Val</td>

<td>Phe</td><td>Leu</td><td>Phe</td><td>Pro</td><td>Pro</td><td>Lys</td><td>Pro</td><td>Lys</td><td>Asp</td><td>Thr</td><td>Leu</td><td>Underworld</td><td>How much</td><td>Cheese</td><td>Arg</td><td>Thr</td>

<td></td><td></td><td></td><td> 260</td><td></td><td></td><td></td><td></td><td> 265</td><td></td><td></td><td></td><td></td><td> 270</td><td></td><td></td>

<td>Pro</td><td>Glu</td><td>Val</td><td>Thr</td><td>Cys</td><td>Val</td><td>val</td><td>Val</td><td>Asp</td><td>Val</td><td>Cheese</td><td>His</td><td>Glu</td><td>Asp</td><td>Pro</td><td>Glu</td>

<td></td><td></td><td> 275</td><td></td><td></td><td></td><td></td><td> 280</td><td></td><td></td><td></td><td></td><td> 285</td><td></td><td></td><td></td>

<td>Val</td><td>Gin</td><td>Phe</td><td>Asn</td><td>Trp</td><td>Tyr</td><td>Val</td><td>Asp</td><td>Gly</td><td>Val</td><td>Glu</td><td>Val</td><td>His</td><td>Asn</td><td>Ala</td><td>Lys</td>

<td></td><td> 290</td><td></td><td></td><td></td><td></td><td> 295</td><td></td><td></td><td></td><td></td><td> 300</td><td></td><td></td><td></td><td></td>

<td>Thr</td><td>Lys</td><td>Pro</td><td>Arg</td><td>Glu</td><td>Glu</td><td>Gin</td><td>Phe</td><td>Asn</td><td>Cheese</td><td>Thr</td><td>Phe</td><td>Arg</td><td>Val</td><td>Val</td><td>Cheese</td>

<td> 305</td><td></td><td></td><td></td><td></td><td> 310</td><td></td><td></td><td></td><td></td><td> 315</td><td></td><td></td><td></td><td></td><td> 320</td>

<td>val</td><td>Leu</td><td>Thr</td><td>Val</td><td>Val</td><td>His</td><td>Gin</td><td>Asp</td><td>Trp</td><td>Leu</td><td>Asn</td><td>Gly</td><td>Lys</td><td>Glu</td><td>Tyr</td><td>lys</td>

<td></td><td></td><td></td><td></td><td> 325</td><td></td><td></td><td></td><td></td><td> 330</td><td></td><td></td><td></td><td></td><td> 335</td><td></td>

<td>Cys</td><td>Lys</td><td>Val</td><td>3 © x</td><td>Asn</td><td>^L.ys</td><td>Gly</td><td>Leu</td><td>Pro</td><td>Ala</td><td>Pro</td><td>How much</td><td>Glu</td><td>Lys</td><td>Thr</td><td>How much</td>

<td></td><td></td><td></td><td> 340</td><td></td><td></td><td></td><td></td><td> 345</td><td></td><td></td><td></td><td></td><td> 350</td><td></td><td></td>

<td>Cheese</td><td>Lys</td><td>Thr</td><td>Lys</td><td>Gly</td><td>Gin</td><td>Pro</td><td>Arg</td><td>Glu</td><td>Pro</td><td>Gin</td><td>Val</td><td>Tyr</td><td></td><td>Leu</td><td>Pro</td>

<td></td><td></td><td> 355</td><td></td><td></td><td></td><td></td><td> 360</td><td></td><td></td><td></td><td></td><td> 365</td><td></td><td></td><td></td>

<td>Pro</td><td>Cheese</td><td>Arg</td><td>Glu</td><td>Glu</td><td>Underworld</td><td>Thr</td><td>Lys</td><td>Asn</td><td>Gin</td><td>Val</td><td>Cheese</td><td>Leu</td><td>Thr</td><td>Cys</td><td>Leu</td>

<td></td><td> 370</td><td></td><td></td><td></td><td></td><td> 375</td><td></td><td></td><td></td><td></td><td> 380</td><td></td><td></td><td></td><td></td>

<td>Val</td><td>Lys</td><td>Gly</td><td>Phe</td><td>Tyr</td><td>Pro</td><td>Cheese</td><td>Asp</td><td>how much</td><td>Ala</td><td>Va.l</td><td>Glu</td><td>Trp</td><td>G1 u</td><td>Cheese</td><td>Asn</td>

<td> 385</td><td></td><td></td><td></td><td></td><td> 390</td><td></td><td></td><td></td><td></td><td> 395</td><td></td><td></td><td></td><td></td><td> 400</td>

<td>Gly</td><td>C-ln</td><td>Pro</td><td>Glu</td><td>Asn</td><td>Asn</td><td>Tyr</td><td>Lys</td><td>Thr</td><td>Thr</td><td>Pro</td><td>Pro</td><td>Underworld</td><td>Leu</td><td>Asp</td><td>Cheese</td>

<td></td><td></td><td></td><td></td><td> 405</td><td></td><td></td><td></td><td></td><td> 410</td><td></td><td></td><td></td><td></td><td> 415</td><td></td>

<td>Asp</td><td>Gly</td><td>Cheese</td><td>Phe</td><td>Phe</td><td>Leu</td><td>Tyr</td><td>Cheese</td><td>Lys</td><td>Leu</td><td>Thr</td><td>Val</td><td>Asp</td><td>Ly p</td><td>Cheese</td><td>Arg</td>

<td></td><td></td><td></td><td> 420</td><td></td><td></td><td></td><td></td><td> 425</td><td></td><td></td><td></td><td></td><td> 430</td><td></td><td></td>

<td>Trp</td><td>Gin</td><td>Gin</td><td>Gly</td><td>Asn</td><td>Val</td><td>Phe</td><td>Cheese</td><td>Cys</td><td>Cheese</td><td>Val</td><td>Underworld</td><td>His</td><td>Glu</td><td>Ara</td><td>Leu</td>

<td></td><td></td><td> 435</td><td></td><td></td><td></td><td></td><td> 440</td><td></td><td></td><td></td><td></td><td> 445</td><td></td><td></td><td></td>

<td>His</td><td>Asn</td><td>His</td><td>Tyr</td><td>Thr</td><td>Gin</td><td>Lys</td><td>Cheese</td><td>Leu</td><td>Cheese</td><td>Leu</td><td>S © K</td><td>Pro</td><td>Gly</td><td>Lys</td><td></td>

450 455 460

<td> &lt;210&gt;</td><td> 64</td>

<td> &lt;211&gt;</td><td> 463</td>

<td> &lt;212&gt;</td><td>PRT</td>

<td> &lt;213&gt;</td><td>Homo sapiens</td>

<td> &lt;400&gt;</td><td> 64</td>

<td>Underworld 1</td><td>Glu</td><td>Phe</td><td>Gly</td><td>Leu 5</td><td>Cheese</td><td colspan="2">Trp Val</td><td>Phe</td><td colspan="2">Leu Val 10</td><td>Ala</td><td>Leu</td><td>Leu</td><td>Arg 15</td><td>Gly</td>

<td>Val</td><td>Gin</td><td>cys</td><td>Gin</td><td>Val</td><td>Gin</td><td>Leu</td><td>Val</td><td>Glu</td><td>Cheese</td><td>Gly</td><td>Giy</td><td>Gly</td><td>Val</td><td>val</td><td>Gin</td>

<td></td><td></td><td></td><td> 20</td><td></td><td></td><td></td><td></td><td> 25</td><td></td><td></td><td></td><td></td><td> 30</td><td></td><td></td>

<td>Pro</td><td>Gly</td><td>Arg</td><td>Cheese</td><td>Leu</td><td>Arg</td><td>Leu</td><td>Cheese</td><td>Cys</td><td>Val</td><td>Ala</td><td>Cheese</td><td>Gly</td><td>Phe</td><td>Thr</td><td>Phe</td>

<td></td><td></td><td> 35</td><td></td><td></td><td></td><td></td><td> 40</td><td></td><td></td><td></td><td></td><td> 45</td><td></td><td></td><td></td>

PL 214 003 Β1

<td>Cheese</td><td>Cheese 50</td><td>His</td><td colspan="2">Gly Met</td><td>His</td><td>Trp 55</td><td colspan="2">Val Arg</td><td>Gin</td><td>Ala</td><td>Pro 60</td><td>Gly</td><td>Lys</td><td>Gly</td><td>Leu</td>

<td>Glu</td><td>Trp</td><td>Val</td><td>Ala</td><td>Val</td><td>How much</td><td>Trp</td><td>Tyr</td><td>Asp</td><td>Gly</td><td>Arg</td><td>Asn</td><td>Lys</td><td>Tyr</td><td>Tyr</td><td>Ala</td>

<td> 65</td><td></td><td></td><td></td><td></td><td> 70</td><td></td><td></td><td></td><td></td><td> 75</td><td></td><td></td><td></td><td></td><td> 80</td>

<td>Asp</td><td>Cheese</td><td>Val</td><td>Lys</td><td>Gly</td><td>Arg</td><td>Phe</td><td>Thr</td><td>How much</td><td>Cheese</td><td>Arg</td><td>Asp</td><td>Asn</td><td>Cheese</td><td>Bald</td><td>Asn</td>

<td></td><td></td><td></td><td></td><td> 85</td><td></td><td></td><td></td><td></td><td> 90</td><td></td><td></td><td></td><td></td><td> 95</td><td></td>

<td>Thr</td><td>Leu</td><td>Phe</td><td>Leu</td><td>Gin</td><td>Underworld</td><td>Asn</td><td>Cheese</td><td>Leu</td><td>Arg</td><td>Ala</td><td>Glu</td><td>Asp</td><td>Thr</td><td>Ala</td><td>Val</td>

<td></td><td></td><td></td><td> 100</td><td></td><td></td><td></td><td></td><td> 105</td><td></td><td></td><td></td><td></td><td> 110</td><td></td><td></td>

<td>Tyr</td><td>Tyr</td><td>cys</td><td>Ala</td><td>Arg</td><td>Gly</td><td>Gly</td><td>His</td><td>Phe</td><td>Gly</td><td>Pro</td><td>Phe</td><td>Asp</td><td>Tyr</td><td>Trp</td><td>Gly</td>

<td></td><td></td><td> 115</td><td></td><td></td><td></td><td></td><td> 120</td><td></td><td></td><td></td><td></td><td> 125</td><td></td><td></td><td></td>

<td>Gin</td><td>Gly</td><td>Thr</td><td>Leu</td><td>Val</td><td>Thr</td><td>Val</td><td>Cheese</td><td>Cheese</td><td>Ala</td><td>Cheese</td><td>Thr</td><td>Lys</td><td>G1 y</td><td>Pro</td><td>Cheese</td>

<td></td><td> 130</td><td></td><td></td><td></td><td></td><td> 135</td><td></td><td></td><td></td><td></td><td> 140</td><td></td><td></td><td></td><td></td>

<td>Val</td><td>Phe</td><td>Pro</td><td>Leu</td><td>Ala</td><td>Pro</td><td>Cys</td><td>Cheese</td><td>Arg</td><td>Cheese</td><td>Thr</td><td>Cheese</td><td>Glu</td><td>Cheese</td><td>Thr</td><td>Ala</td>

<td> 145</td><td></td><td></td><td></td><td></td><td> 150</td><td></td><td></td><td></td><td></td><td> 155</td><td></td><td></td><td></td><td></td><td> 160</td>

<td>Ala</td><td>Leu</td><td>Gly</td><td>Cys</td><td>Leu</td><td>Val</td><td>Lys</td><td>Asp</td><td>Tyr</td><td>Phe</td><td>Pro</td><td>Glu</td><td>Pro</td><td>Val</td><td>Thr</td><td>Val</td>

<td></td><td></td><td></td><td></td><td> 165</td><td></td><td></td><td></td><td></td><td> 170</td><td></td><td></td><td></td><td></td><td> 175</td><td></td>

<td>Cheese</td><td>Trp</td><td>Asn</td><td>Cheese</td><td>Gly</td><td>Ala</td><td>Leu</td><td>Thr</td><td>Cheese</td><td>Gly</td><td>Val</td><td>His</td><td>Thr</td><td>Phe</td><td>Pro</td><td>Ala</td>

<td></td><td></td><td></td><td> 180</td><td></td><td></td><td></td><td></td><td> 185</td><td></td><td></td><td></td><td></td><td> 190</td><td></td><td></td>

<td>Va 1</td><td>Leu</td><td>Gin</td><td>Cheese</td><td>Cheese</td><td>Gly</td><td>Leu</td><td>Tyr</td><td>Cheese</td><td>Leu</td><td>Cheese</td><td>Cheese</td><td>Val</td><td>Val</td><td>Thr</td><td>Val</td>

<td></td><td></td><td> 195</td><td></td><td></td><td></td><td></td><td> 200</td><td></td><td></td><td></td><td></td><td> 205</td><td></td><td></td><td></td>

<td>Pro</td><td>Cheese</td><td>Cheese</td><td>Asn</td><td>Phe</td><td>Gly</td><td>Thr</td><td>Gin</td><td>Thr</td><td>Tyr</td><td>Thr</td><td>cys</td><td>Asn</td><td>Val</td><td>Asp</td><td>His</td>

<td></td><td> 210</td><td></td><td></td><td></td><td></td><td> 215</td><td></td><td></td><td></td><td></td><td> 220</td><td></td><td></td><td></td><td></td>

<td>Lys</td><td>Pro</td><td>Cheese</td><td>Asn</td><td>Thr</td><td>Lys</td><td>Va 1</td><td>Asp</td><td>Lys</td><td>Thr</td><td>Val</td><td>Glu</td><td>Arg</td><td>Lys</td><td>Cys</td><td>Cys</td>

<td> 225</td><td></td><td></td><td></td><td></td><td> 230</td><td></td><td></td><td></td><td></td><td> 235</td><td></td><td></td><td></td><td></td><td> 240</td>

<td>Val</td><td>Glu</td><td>Cys</td><td>Pro</td><td>Pro</td><td>Cys</td><td>Pro</td><td>Ala</td><td>Pro</td><td>Pro</td><td>Val</td><td>Ala</td><td>Gly</td><td>Pro</td><td>Cheese</td><td>Val</td>

<td></td><td></td><td></td><td></td><td> 245</td><td></td><td></td><td></td><td></td><td> 250</td><td></td><td></td><td></td><td></td><td> 255</td><td></td>

<td>Phe</td><td>Leu</td><td>Phe</td><td>Pro</td><td>Pro</td><td>Lys</td><td>Pro</td><td>Lys</td><td>Asp</td><td>Thr</td><td>Leu</td><td>Underworld</td><td>How much</td><td>Cheese</td><td>Arg</td><td>Thr</td>

<td></td><td></td><td></td><td> 260</td><td></td><td></td><td></td><td></td><td> 265</td><td></td><td></td><td></td><td></td><td> 270</td><td></td><td></td>

<td>Pro</td><td>Glu</td><td>Val</td><td>Thr</td><td>Cys</td><td>Val</td><td>Val</td><td>Val</td><td>Asp</td><td>Val</td><td>Cheese</td><td>His</td><td>Glu</td><td>Asp</td><td>Pro</td><td>Glu</td>

<td></td><td></td><td> 275</td><td></td><td></td><td></td><td></td><td> 280</td><td></td><td></td><td></td><td></td><td> 285</td><td></td><td></td><td></td>

<td>Val</td><td>Gin</td><td>Phe</td><td>Asn</td><td>Trp</td><td>Tyr</td><td>Val</td><td>Asp</td><td>Gly</td><td>val</td><td>Glu</td><td>Val</td><td>His</td><td>Asn</td><td>Ala</td><td>Lys</td>

<td></td><td> 290</td><td></td><td></td><td></td><td></td><td> 295</td><td></td><td></td><td></td><td></td><td> 300</td><td></td><td></td><td></td><td></td>

<td>Thr</td><td>Lys</td><td>Pro</td><td>Arg</td><td>Glu</td><td>Glu</td><td>Gin</td><td>Phe</td><td>Gin</td><td>Cheese</td><td>Thr</td><td>Phe</td><td>Arg</td><td>Val</td><td>val</td><td>Cheese</td>

<td> 305</td><td></td><td></td><td></td><td></td><td> 310</td><td></td><td></td><td></td><td></td><td> 315</td><td></td><td></td><td></td><td></td><td> 320</td>

<td>Val</td><td>Leu</td><td>Thr</td><td>Val</td><td>Val</td><td>His</td><td>Gin</td><td>Asp</td><td>Trp</td><td>Leu</td><td>Asn</td><td>dy</td><td>Lys</td><td>Glu</td><td>ψγ y-</td><td>Lys</td>

<td></td><td></td><td></td><td></td><td> 325</td><td></td><td></td><td></td><td></td><td> 330</td><td></td><td></td><td></td><td></td><td> 335</td><td></td>

<td>Cys</td><td>Lys</td><td>Val</td><td>Cheese</td><td>Asn</td><td>Lys</td><td>Gly</td><td>Ii eu</td><td>Pro</td><td>Ala</td><td>Pro</td><td>How much</td><td>Glu</td><td>Lys</td><td>Thr</td><td>How much</td>

<td></td><td></td><td></td><td> 340</td><td></td><td></td><td></td><td></td><td> 345</td><td></td><td></td><td></td><td></td><td> 350</td><td></td><td></td>

PL 214 003 Β1

<td>Cheese</td><td>Lys</td><td>Thr 355</td><td>Lys</td><td>Gly</td><td>Gin</td><td>Pro</td><td>Arg 360</td><td>Glu</td><td>Pro</td><td>Gin</td><td>Val</td><td>Tyr 365</td><td>Thr</td><td>Leu</td><td>Pro</td>

<td>Pro</td><td>Cheese</td><td>Arg</td><td>Glu</td><td>Glu</td><td>Underworld</td><td>Thr</td><td>Lys</td><td>Asn</td><td>Gin</td><td>Val</td><td>Cheese</td><td>Leu</td><td>Thr</td><td>Cys</td><td>Leu</td>

<td></td><td> 370</td><td></td><td></td><td></td><td></td><td> 375</td><td></td><td></td><td></td><td></td><td> 380</td><td></td><td></td><td></td><td></td>

<td>vai</td><td>Lys</td><td>Gly</td><td>Phe</td><td>Tyr</td><td>Pro</td><td>Cheese</td><td>Asp</td><td>How much</td><td>Ala</td><td>Val</td><td>Glu</td><td>Trp</td><td>Glu</td><td>Cheese</td><td>Asn</td>

<td> 385</td><td></td><td></td><td></td><td></td><td> 390</td><td></td><td></td><td></td><td></td><td> 395</td><td></td><td></td><td></td><td></td><td> 400</td>

<td>Gly</td><td>Gin</td><td>Pro</td><td>Glu</td><td>Asn</td><td>Asn</td><td>Tyr</td><td>Lys</td><td>Thr</td><td>Thr</td><td>Pro</td><td>Pro</td><td>Underworld</td><td>Leu</td><td>Asp</td><td>Cheese</td>

<td></td><td></td><td></td><td></td><td> 405</td><td></td><td></td><td></td><td></td><td> 410</td><td></td><td></td><td></td><td></td><td> 415</td><td></td>

<td>Asp</td><td>Gly</td><td>Cheese</td><td>Phe</td><td>Phe</td><td>Leu</td><td>Tyr</td><td>Cheese</td><td>Lys</td><td>Leu</td><td>Thr</td><td>Val</td><td>Asp</td><td>Lys</td><td>Cheese</td><td>Arg</td>

<td></td><td></td><td></td><td> 420</td><td></td><td></td><td></td><td></td><td> 425</td><td></td><td></td><td></td><td></td><td> 430</td><td></td><td></td>

<td>Trp</td><td>G1 n</td><td>Gin</td><td>Gly</td><td>Asn</td><td>Val</td><td>Phe</td><td>Cheese</td><td>cys</td><td>Cheese</td><td>y ci 1.</td><td>Underworld</td><td>His</td><td>G1 u</td><td>Ala</td><td>Leu</td>

<td></td><td></td><td> 435</td><td></td><td></td><td></td><td></td><td> 440</td><td></td><td></td><td></td><td></td><td> 445</td><td></td><td></td><td></td>

<td>His</td><td>Asn</td><td>His</td><td>Tyr</td><td>Thr</td><td>Gin</td><td>Lys</td><td>Cheese</td><td>Leu</td><td>Cheese</td><td>Leu</td><td>Cheese</td><td>Pro</td><td>ciy</td><td>Lys</td><td></td>

450 455 460 &lt;210&gt; 65 &lt;211&gt; 235 &lt;212&gt; PRT &lt;213&gt; Homo sapiens &lt;400&gt; 65

<td>Underworld</td><td>Glu</td><td>Thr</td><td>Pro</td><td>Ala 5</td><td>Gin</td><td>Leu</td><td>Leu</td><td>Phe</td><td>Leu 10</td><td>Leu</td><td>Leu</td><td>Leu</td><td>Trp</td><td>Leu 15</td><td>Pro</td>

<td>Asp</td><td>Thr</td><td>Thr</td><td>Gly</td><td>Glu</td><td>How much</td><td>Val</td><td>Leu</td><td>Thr</td><td>Gin</td><td>Cheese</td><td>Pro</td><td>Gly</td><td>Thr</td><td>Leu</td><td>Cheese</td>

<td></td><td></td><td></td><td> 20</td><td></td><td></td><td></td><td></td><td> 25</td><td></td><td></td><td></td><td></td><td> 30</td><td></td><td></td>

<td>Leu</td><td>Cheese</td><td>Pro</td><td>Gly</td><td>Glu</td><td>Arg</td><td>Ala</td><td>Thr</td><td>Leu</td><td>Cheese</td><td>Cys</td><td>Arg</td><td>Ala</td><td>Cheese</td><td>Gin</td><td>Cheese</td>

<td></td><td></td><td> 35</td><td></td><td></td><td></td><td></td><td> 40</td><td></td><td></td><td></td><td></td><td> 45</td><td></td><td></td><td></td>

<td>How much</td><td>Cheese</td><td>Cheese</td><td>Cheese</td><td>Phe</td><td>Leu</td><td>Ala</td><td>Trp</td><td>Tyr</td><td>Gin</td><td>Gin</td><td>Arg</td><td>Pro</td><td>Gly</td><td>Gin</td><td>Ala</td>

<td></td><td> 50</td><td></td><td></td><td></td><td></td><td> 55</td><td></td><td></td><td></td><td></td><td> 60</td><td></td><td></td><td></td><td></td>

<td>Pro</td><td>Arg</td><td>Leu</td><td>Leu</td><td>How much</td><td>Tyr</td><td>Gly</td><td>Ala</td><td>Cheese</td><td>Cheese</td><td>Arg</td><td>Ala</td><td>Thr</td><td>Gly</td><td>how much</td><td>Pro</td>

<td> 65</td><td></td><td></td><td></td><td></td><td> 70</td><td></td><td></td><td></td><td></td><td> 75</td><td></td><td></td><td></td><td></td><td> 80</td>

<td>Asp</td><td>Arg</td><td>Phe</td><td>Cheese</td><td>Gly</td><td>Cheese</td><td>Gly</td><td>Cheese</td><td>Gly</td><td>Thr</td><td>Asp</td><td>Phe</td><td>Thr</td><td>Leu</td><td>Thr</td><td>How much</td>

<td></td><td></td><td></td><td></td><td> 85</td><td></td><td></td><td></td><td></td><td> 90</td><td></td><td></td><td></td><td></td><td> 95</td><td></td>

<td>Cheese</td><td>Arg</td><td>Leu</td><td>Glu</td><td>Pro</td><td>Glu</td><td>Asp</td><td>Phe</td><td>Ala</td><td>Val</td><td>Tyr</td><td>Tyr</td><td>Cys</td><td>Gin.</td><td>Gin</td><td>Tyr</td>

<td></td><td></td><td></td><td> 100</td><td></td><td></td><td></td><td></td><td> 105</td><td></td><td></td><td></td><td></td><td> 110</td><td></td><td></td>

<td>Gly</td><td>Thr</td><td>Cheese</td><td>Pro</td><td>Trp</td><td>Thr</td><td>Phe</td><td>Gly</td><td>Gin</td><td>Gly</td><td>Thr</td><td>Lys</td><td>Val</td><td>Glu</td><td>How much</td><td>Lys</td>

<td></td><td></td><td> 115</td><td></td><td></td><td></td><td></td><td> 120</td><td></td><td></td><td></td><td></td><td> 125</td><td></td><td></td><td></td>

<td>Arg</td><td>Thr</td><td>vai</td><td>Ala</td><td>Ala</td><td>Pro</td><td>Cheese</td><td>Val</td><td>Phe</td><td>How much</td><td>Phe</td><td>Pro</td><td>Pro</td><td>Cheese</td><td>Asp</td><td>Glu</td>

<td></td><td> 130</td><td></td><td></td><td></td><td></td><td> 135</td><td></td><td></td><td></td><td></td><td> 140</td><td></td><td></td><td></td><td></td>

<td>Gin</td><td>Leu</td><td>Lys</td><td>Cheese</td><td>Gly</td><td>Thr</td><td>Ala</td><td>Cheese</td><td>Val</td><td>Val</td><td>Cys</td><td>Leu</td><td>Leu</td><td>Asn</td><td>Asn</td><td>Phe</td>

145 150 155 160

PL 214 003 Β1

<td>Tyr</td><td>Pro</td><td>Arg</td><td>Glu</td><td>Ala 165</td><td>Lys</td><td>Val</td><td>Gin</td><td>Trp</td><td>Lys 170</td><td>Val</td><td>Asp</td><td>Asn</td><td>Al a</td><td>Leu 175</td><td>Gin</td>

<td>Cheese</td><td>Gly</td><td>Asn</td><td>Cheese</td><td>Gin</td><td>Glu</td><td>Cheese</td><td>Val</td><td>Thr</td><td>Glu</td><td>Gin</td><td>Asp</td><td>Cheese</td><td>Lys</td><td>Asp</td><td>Cheese</td>

<td></td><td></td><td></td><td> 180</td><td></td><td></td><td></td><td></td><td> 185</td><td></td><td></td><td></td><td></td><td> 190</td><td></td><td></td>

<td>Thr</td><td>Tyr</td><td>Cheese</td><td>Leu</td><td>Cheese</td><td>Cheese</td><td>Thr</td><td>Leu</td><td>Thr</td><td>Leu</td><td>Cheese</td><td>Lys</td><td>Ala</td><td>Asp</td><td>Tyr</td><td>Glu</td>

<td></td><td></td><td> 195</td><td></td><td></td><td></td><td></td><td> 200</td><td></td><td></td><td></td><td></td><td> 205</td><td></td><td></td><td></td>

<td>Lys</td><td>His</td><td>Lys</td><td>Val</td><td>Tyr</td><td>Ala</td><td>Cys</td><td>Glu</td><td>Val</td><td>Thr</td><td>His</td><td>Gin</td><td>Gly</td><td>Leu</td><td>Cheese</td><td>Cheese</td>

<td></td><td> 210</td><td></td><td></td><td></td><td></td><td> 215</td><td></td><td></td><td></td><td></td><td> 220</td><td></td><td></td><td></td><td></td>

<td>Pro</td><td>Val</td><td>Thr</td><td>Lys</td><td>Cheese</td><td>Phe</td><td>Asn</td><td>Arg</td><td>Gly</td><td>Glu</td><td>Cys</td><td></td><td></td><td></td><td></td><td></td>

225 230 235 &lt;210 66 &lt;211&gt; 464 &lt;212&gt; PRT &lt;213&gt; Homo sapiens

<td colspan="3"> &lt;400 66</td><td colspan="2" rowspan="2">Gly Leu 5</td><td colspan="3" rowspan="2">Trp Val cheese</td><td rowspan="2">Phe</td><td colspan="2" rowspan="2">Leu Val 10</td><td rowspan="2">Ala</td><td rowspan="2">Leu</td><td rowspan="2">Leu</td><td rowspan="2">Arg 15</td><td rowspan="2">Gly</td>

<td>Underworld 1</td><td>Glu</td><td>Phe</td>

<td>Val</td><td>Gin</td><td>Cys</td><td>Gin</td><td>Val</td><td>Gin</td><td>Leu</td><td>val</td><td>Glu</td><td>Cheese</td><td>Gly</td><td>Gly</td><td>Gly</td><td>Val</td><td>Val</td><td>Gin</td>

<td></td><td></td><td></td><td> 20</td><td></td><td></td><td></td><td></td><td> 25</td><td></td><td></td><td></td><td></td><td> 30</td><td></td><td></td>

<td>Ero</td><td>Gly</td><td>Arg</td><td>Cheese</td><td>Leu</td><td>Arg</td><td>Leu</td><td>Cheese</td><td>Cys</td><td>Thr</td><td>Ala</td><td>Cheese</td><td>Giy</td><td>Phe</td><td>Thr</td><td>Phe</td>

<td></td><td></td><td> 35</td><td></td><td></td><td></td><td></td><td> 40</td><td></td><td></td><td></td><td></td><td> 45</td><td></td><td></td><td></td>

<td>Cheese</td><td>Asn</td><td>Tyr</td><td>Gly</td><td>Underworld</td><td>His</td><td>Trp</td><td>val</td><td>Arg</td><td>Gin</td><td>Ala</td><td>Pro</td><td>Gly</td><td>Lys</td><td>Gly</td><td>Leu</td>

<td></td><td> 50</td><td></td><td></td><td></td><td></td><td> 55</td><td></td><td></td><td></td><td></td><td> 60</td><td></td><td></td><td></td><td></td>

<td>Glu</td><td>Trp</td><td>Val</td><td>Ala</td><td>Val</td><td>How much</td><td>Trp</td><td>Tyr</td><td>Asp</td><td>Gly</td><td>Cheese</td><td>Asn</td><td>Lys</td><td>His</td><td>Tyr</td><td>Gly</td>

<td> 65</td><td></td><td></td><td></td><td></td><td> 70</td><td></td><td></td><td></td><td></td><td> 75</td><td></td><td></td><td></td><td></td><td> 80</td>

<td>Asp</td><td>5 OX '</td><td>Val</td><td>Lys</td><td>Gly</td><td>Arg</td><td>Phe</td><td>Thr</td><td>How much</td><td>Cheese</td><td>Cheese</td><td>Asp</td><td>Asn</td><td>Cheese</td><td>Lys</td><td>Asn</td>

<td></td><td></td><td></td><td></td><td> 85</td><td></td><td></td><td></td><td></td><td> 90</td><td></td><td></td><td></td><td></td><td> 95</td><td></td>

<td>Thr</td><td>Leu</td><td>Tyr</td><td>Leu</td><td>Gin</td><td>Underworld</td><td>Asn</td><td>Cheese</td><td>^ 3 * H</td><td>Arg</td><td>Ala</td><td>Glu</td><td>Asp</td><td>Thr</td><td>Ala</td><td>Val</td>

<td></td><td></td><td></td><td> 100</td><td></td><td></td><td></td><td></td><td> 105</td><td></td><td></td><td></td><td></td><td> 110</td><td></td><td></td>

<td>Tyr</td><td>Tyr</td><td>Cys</td><td>Ala</td><td>Arg</td><td>Gly</td><td>Glu</td><td>Arg</td><td>Leu</td><td>Gly</td><td>Cheese</td><td>Tyr</td><td>Phe</td><td>Asp</td><td>Tyr</td><td>Trp</td>

<td></td><td></td><td> 115</td><td></td><td></td><td></td><td></td><td> 120</td><td></td><td></td><td></td><td></td><td> 125</td><td></td><td></td><td></td>

<td>Gly</td><td>Gin</td><td>Gly</td><td>Thr</td><td>Leu</td><td>Val</td><td>Thr</td><td>Val</td><td>Cheese</td><td>Cheese</td><td>Ala</td><td>Cheese</td><td>Thr</td><td>Lys</td><td>Gly</td><td>Pro</td>

<td></td><td> 130</td><td></td><td></td><td></td><td></td><td> 135</td><td></td><td></td><td></td><td></td><td> 140</td><td></td><td></td><td></td><td></td>

<td>Cheese</td><td>Val</td><td>Phe</td><td>Pro</td><td>Leu</td><td>Ala</td><td>Pro</td><td>Cys</td><td>cheese</td><td>Arg</td><td>Cheese</td><td>Thr</td><td>Cheese</td><td>Glu</td><td>Cheese</td><td>Thr</td>

<td> 145</td><td></td><td></td><td></td><td></td><td> 150</td><td></td><td></td><td></td><td></td><td> 155</td><td></td><td></td><td></td><td></td><td> 160</td>

<td>Ala</td><td>Ala</td><td>Leu</td><td>Gly</td><td>Cys</td><td>Leu</td><td>Val</td><td>Lys</td><td>Asp</td><td>Tyr</td><td>Phe</td><td>Pro</td><td>Glu</td><td>Pro</td><td>Val</td><td>Thr</td>

<td></td><td></td><td></td><td></td><td> 165</td><td></td><td></td><td></td><td></td><td> 170</td><td></td><td></td><td></td><td></td><td> 175</td><td></td>

<td>Val</td><td>Cheese</td><td>Trp</td><td>Asn</td><td>Cheese</td><td>Gly</td><td>Ala</td><td>Leu</td><td>Thr</td><td>Cheese</td><td>dy</td><td>Val</td><td>His</td><td>Thr</td><td>Phe</td><td>Pro</td>

<td></td><td></td><td></td><td> 180</td><td></td><td></td><td></td><td></td><td> 185</td><td></td><td></td><td></td><td></td><td> 190</td><td></td><td></td>

PL 214 003 Β1

<td>Ala</td><td>Val</td><td>Leu 195</td><td>Gin</td><td>Cheese</td><td>Cheese</td><td>Gly</td><td>Leu 200</td><td>Tyr</td><td>Se r</td><td>Leu</td><td>Cheese</td><td>Cheese 205</td><td>Val.</td><td>Val</td><td>Thr</td>

<td>Val</td><td>Pro</td><td>Cheese</td><td>Cheese</td><td>Asn</td><td>Phe</td><td>Gly</td><td>Thr</td><td>Gin</td><td>Thr</td><td>Tyr</td><td>Thr</td><td>Cys</td><td>Asn</td><td>Val</td><td>Asp</td>

<td></td><td> 210</td><td></td><td></td><td></td><td></td><td> 215</td><td></td><td></td><td></td><td></td><td> 220</td><td></td><td></td><td></td><td></td>

<td>His</td><td>Lys</td><td>Pro</td><td>Cheese</td><td>Asn</td><td>Thr</td><td>Lys</td><td>Val</td><td>Asp</td><td>Lys</td><td>Thr</td><td>Val</td><td>Glu</td><td>Arg</td><td>Lys</td><td>cys</td>

<td> 225</td><td></td><td></td><td></td><td></td><td> 230</td><td></td><td></td><td></td><td></td><td> 235</td><td></td><td></td><td></td><td></td><td> 240</td>

<td>Cys</td><td>Val</td><td>Glu</td><td>Cys</td><td>Pro</td><td>Pro</td><td>Cys</td><td>Pro</td><td>Ala</td><td>Pro</td><td>Pro</td><td>Val</td><td>Ala</td><td>Gly</td><td>Pro</td><td>Cheese</td>

<td></td><td></td><td></td><td></td><td> 245</td><td></td><td></td><td></td><td></td><td> 250</td><td></td><td></td><td></td><td></td><td> 255</td><td></td>

<td>Val</td><td>Phe</td><td>Leu</td><td>Phe</td><td>Pro</td><td>Pro</td><td>Lys</td><td>Pro</td><td>Lys</td><td>Asp</td><td>Thr</td><td>Leu</td><td>Underworld</td><td>How much</td><td>Cheese</td><td>Arg</td>

<td></td><td></td><td></td><td> 260</td><td></td><td></td><td></td><td></td><td> 2 65</td><td></td><td></td><td></td><td></td><td> 270</td><td></td><td></td>

<td>Thr</td><td>Pro</td><td>Glu</td><td>Val</td><td>Thr</td><td>Cys</td><td>Val</td><td>Val</td><td>Val</td><td>Asp</td><td>Val</td><td>Cheese</td><td>His</td><td>Glu</td><td>Asp</td><td>Pro</td>

<td></td><td></td><td> 275</td><td></td><td></td><td></td><td></td><td> 280</td><td></td><td></td><td></td><td></td><td> 285</td><td></td><td></td><td></td>

<td>Glu</td><td>Val</td><td>Gin</td><td>Phe</td><td>Asn</td><td>Trp</td><td>Tyr</td><td>Val</td><td>Asp</td><td>Gly</td><td>Val</td><td>Glu</td><td>Val</td><td>His</td><td>Asn</td><td>Ala</td>

<td></td><td> 290</td><td></td><td></td><td></td><td></td><td> 295</td><td></td><td></td><td></td><td></td><td> 300</td><td></td><td></td><td></td><td></td>

<td>Lys</td><td>Thr</td><td>Lys</td><td>Pro</td><td>Arg</td><td>Glu</td><td>Glu</td><td>Gin</td><td>Phe</td><td>Asn</td><td>Cheese</td><td>Thr</td><td>Phe</td><td>Arg</td><td>Val</td><td>Val</td>

<td> 305</td><td></td><td></td><td></td><td></td><td> 310</td><td></td><td></td><td></td><td></td><td> 315</td><td></td><td></td><td></td><td></td><td> 320</td>

<td>Cheese</td><td>Val</td><td>Leu</td><td>Thr</td><td>Val</td><td>Val</td><td>His</td><td>Gin</td><td>Asp</td><td>Trp</td><td>Leu</td><td>Asn</td><td>Gly</td><td>Lys</td><td>Glu</td><td>Tyr</td>

<td></td><td></td><td></td><td></td><td> 325</td><td></td><td></td><td></td><td></td><td> 330</td><td></td><td></td><td></td><td></td><td> 335</td><td></td>

<td>Lys</td><td>Cys</td><td>Lys</td><td>Val</td><td>Cheese</td><td>Asn</td><td>Lys</td><td>Gly</td><td>Leu</td><td>Pro</td><td>Ala</td><td>Pro</td><td>How much</td><td>Glu</td><td>Lys</td><td>Thr</td>

<td></td><td></td><td></td><td> 340</td><td></td><td></td><td></td><td></td><td> 345</td><td></td><td></td><td></td><td></td><td> 350</td><td></td><td></td>

<td>Background</td><td>Cheese</td><td>Lys</td><td>Thr</td><td>Lys</td><td>Gly</td><td>Gin</td><td>Pro</td><td>Arg</td><td>Glu</td><td>Pro</td><td>Gin</td><td>Val</td><td>Tyr</td><td>Thr</td><td>Leu</td>

<td></td><td></td><td> 355</td><td></td><td></td><td></td><td></td><td> 360</td><td></td><td></td><td></td><td></td><td> 365</td><td></td><td></td><td></td>

<td>Pro</td><td>Pro</td><td>Cheese</td><td>Arg</td><td>Glu</td><td>Glu</td><td>Underworld</td><td>Thr</td><td>Lys</td><td>Asn</td><td>Gin</td><td>Val</td><td>Cheese</td><td>Leu</td><td>Thr</td><td>Cys</td>

<td></td><td> 370</td><td></td><td></td><td></td><td></td><td> 375</td><td></td><td></td><td></td><td></td><td> 380</td><td></td><td></td><td></td><td></td>

<td>Leu</td><td>Val</td><td>Lys</td><td>Gly</td><td>Phe</td><td>Tyr</td><td>Pro</td><td>Cheese</td><td>Asp</td><td>How much</td><td>Ala</td><td>Val</td><td>Glu</td><td>Trp</td><td>Glu</td><td>Cheese</td>

<td> 385</td><td></td><td></td><td></td><td></td><td> 390</td><td></td><td></td><td></td><td></td><td> 395</td><td></td><td></td><td></td><td></td><td> 400</td>

<td>Asn</td><td>Gly</td><td>Gin</td><td>Pro</td><td>Glu</td><td>Asn</td><td>Asn</td><td>Tyr</td><td>Lys</td><td>Thr</td><td>Thr</td><td>Pro</td><td>Pro</td><td>Underworld</td><td>L ίί</td><td>Asp</td>

<td></td><td></td><td></td><td></td><td> 405</td><td></td><td></td><td></td><td></td><td> 410</td><td></td><td></td><td></td><td></td><td> 415</td><td></td>

<td>Cheese</td><td>Asp</td><td>Gly</td><td>Cheese</td><td>Phe</td><td>Phe</td><td>Leu</td><td>Tyr</td><td>Cheese</td><td>Lys</td><td>Leu</td><td>Thr</td><td>Val</td><td>Asp</td><td>Lys</td><td>Cheese</td>

<td></td><td></td><td></td><td> 420</td><td></td><td></td><td></td><td></td><td> 425</td><td></td><td></td><td></td><td></td><td> 430</td><td></td><td></td>

<td>Arg</td><td>Trp</td><td>Gin</td><td>Gin</td><td>Gly</td><td>Asn</td><td>Val</td><td>Phe</td><td>Cheese</td><td>cys</td><td>Cheese</td><td>Val</td><td>Underworld</td><td>His</td><td>Glu</td><td>Ala</td>

<td></td><td></td><td> 435</td><td></td><td></td><td></td><td></td><td> 440</td><td></td><td></td><td></td><td></td><td> 445</td><td></td><td></td><td></td>

<td>Leu</td><td>His</td><td>Asn</td><td>His</td><td>Tyr</td><td>Thr</td><td>Gin</td><td>Lys</td><td>Cheese</td><td>Leu</td><td>Cheese</td><td>Leu</td><td>Cheese</td><td>Pro</td><td>Gly</td><td>Lys</td>

<td></td><td> 450</td><td></td><td></td><td></td><td></td><td> 455</td><td></td><td></td><td></td><td></td><td> 460</td><td></td><td></td><td></td><td></td>

&lt;210&gt; 67 &lt;211&gt; 233 &lt;2121 PRT &lt;213&gt; Homo sapiens &lt;400 67

PL 214 003 Β1

<td>Underworld 1</td><td>Glu</td><td>Thr</td><td>Pro</td><td>L, ci 5</td><td>Gin</td><td>Leu</td><td>Leu</td><td>Phe</td><td>Leu</td><td>Leu</td><td>Leu</td><td>Leu</td><td>Trp</td><td>Leu 15</td><td>Pro</td>

<td>Asp</td><td>Thr</td><td>Thr</td><td>dy</td><td>G1 u</td><td>How much</td><td>Val</td><td>Leu</td><td>Thr</td><td>Gin</td><td>Cheese</td><td>Pro</td><td>Gly</td><td>Thr</td><td>Leu</td><td>Cheese</td>

<td></td><td></td><td></td><td> 20</td><td></td><td></td><td></td><td></td><td>'“Ic Z. □</td><td></td><td></td><td></td><td></td><td> 30</td><td></td><td></td>

<td>Leu</td><td>Cheese</td><td>Pro</td><td>Gly</td><td>Glu</td><td>Arg</td><td>Ala</td><td>Thr</td><td>Leu</td><td>Cheese</td><td>Cys</td><td>Arg</td><td>Thr</td><td>Cheese</td><td>Val</td><td>Cheese</td>

<td></td><td></td><td> 35</td><td></td><td></td><td></td><td></td><td> 40</td><td></td><td></td><td></td><td></td><td> 45</td><td></td><td></td><td></td>

<td>Cheese</td><td>Cheese</td><td>Tyr</td><td>Leu</td><td>Ala</td><td>Trp</td><td>Tyr</td><td>Gin</td><td>Gin</td><td>Lys</td><td>Pro</td><td>Gly</td><td>Gin</td><td>Ala</td><td>Pro</td><td>Arg</td>

<td></td><td> 50</td><td></td><td></td><td></td><td></td><td> 55</td><td></td><td></td><td></td><td></td><td> 60</td><td></td><td></td><td></td><td></td>

<td>Leu</td><td>Leu</td><td>How much</td><td>Tyr</td><td>Gly</td><td>Ala</td><td>Cheese</td><td>Cheese</td><td>Arg</td><td>Ala</td><td>Thr</td><td>Gly</td><td>How much</td><td>Pro</td><td>Asp</td><td>Arg</td>

<td> 65</td><td></td><td></td><td></td><td></td><td> 70</td><td></td><td></td><td></td><td></td><td> 75</td><td></td><td></td><td></td><td></td><td> 80</td>

<td>Phe</td><td>Cheese</td><td>strength</td><td>Cheese</td><td>G1 y</td><td>Cheese</td><td>Gly</td><td>Thr</td><td>Asp</td><td>Phe</td><td>Thr</td><td>Leu</td><td>Thr</td><td>How much</td><td>Cheese</td><td>Arg</td>

<td></td><td></td><td></td><td></td><td> 8 5</td><td></td><td></td><td></td><td></td><td> 90</td><td></td><td></td><td></td><td></td><td> 95</td><td></td>

<td>Leu</td><td>Glu</td><td>Pro</td><td>Glu</td><td>Asp</td><td>Phe</td><td>Ala</td><td>Val</td><td>Tyr</td><td>Tyr</td><td>Cys</td><td>Gin</td><td>Gin</td><td>Tyr</td><td>Gly</td><td>How much</td>

<td></td><td></td><td></td><td> 100</td><td></td><td></td><td></td><td></td><td> 105</td><td></td><td></td><td></td><td></td><td> 110</td><td></td><td></td>

<td>Cheese</td><td>Pro</td><td>Phe</td><td>Thr</td><td>Phe</td><td>Gly</td><td>Gly</td><td>Gly</td><td>Thr</td><td>Lys</td><td>Val</td><td>Glu</td><td>How much</td><td>Lys</td><td>Arg</td><td>Thr</td>

<td></td><td></td><td> 115</td><td></td><td></td><td></td><td></td><td> 120</td><td></td><td></td><td></td><td></td><td> 125</td><td></td><td></td><td></td>

<td>Val</td><td>Ala</td><td>Ala</td><td>Pro</td><td>Cheese</td><td>val</td><td>Phe</td><td>How much</td><td>Phe</td><td>Pro</td><td>Pro</td><td>Cheese</td><td>Asp</td><td>Glu</td><td>Gin</td><td>Leu</td>

<td></td><td> 130</td><td></td><td></td><td></td><td></td><td> 135</td><td></td><td></td><td></td><td></td><td> 140</td><td></td><td></td><td></td><td></td>

<td>Lys</td><td>Cheese</td><td>Gly</td><td>Thr</td><td>Ala</td><td>Cheese</td><td>Val</td><td>Val</td><td>cys</td><td>Leu</td><td>Leu</td><td>Asn</td><td>Asn</td><td>Phe</td><td>Tyr</td><td>Pro</td>

<td> 145</td><td></td><td></td><td></td><td></td><td> 150</td><td></td><td></td><td></td><td></td><td> 155</td><td></td><td></td><td></td><td></td><td> 160</td>

<td>Arg</td><td>Glu</td><td>Ala</td><td>Lys</td><td>Val</td><td>Gin</td><td>Trp</td><td>Lys</td><td>Val</td><td>Asp</td><td>Asn</td><td>Ala</td><td>Leu</td><td>Gin</td><td>Cheese</td><td>Gly</td>

<td></td><td></td><td></td><td></td><td> 165</td><td></td><td></td><td></td><td></td><td> 170</td><td></td><td></td><td></td><td></td><td> 175</td><td></td>

<td>Asn</td><td>Cheese</td><td>G1 n</td><td>Glu</td><td>Cheese</td><td>Val</td><td>Thr</td><td>Glu</td><td>Gin</td><td>Asp</td><td>Cheese</td><td>Lys</td><td>Asp</td><td>Cheese</td><td>Thr</td><td>Tyr</td>

<td></td><td></td><td></td><td> 18 0</td><td></td><td></td><td></td><td></td><td> 185</td><td></td><td></td><td></td><td></td><td> 190</td><td></td><td></td>

<td>Cheese</td><td>Leu</td><td>Cheese</td><td>Cheese</td><td>Thr</td><td>Leu</td><td>Thr</td><td>Leu</td><td>Cheese</td><td>Lys</td><td>Ala</td><td>Asp</td><td>Tyr</td><td>Glu</td><td>Lys</td><td>His</td>

<td></td><td></td><td> 195</td><td></td><td></td><td></td><td></td><td> 200</td><td></td><td></td><td></td><td></td><td> 205</td><td></td><td></td><td></td>

<td>Lys</td><td>Val</td><td>Tyr</td><td>Al 3</td><td>cys</td><td>Glu</td><td>val</td><td>Thr</td><td>His</td><td>Gin</td><td>Gly</td><td>Lbu</td><td>Cheese</td><td>Cheese</td><td>Pro</td><td>Val</td>

<td></td><td> 210</td><td></td><td></td><td></td><td></td><td> 215</td><td></td><td></td><td></td><td></td><td> 220</td><td></td><td></td><td></td><td></td>

<td>Thr</td><td>Bald</td><td>Cheese</td><td>Phe</td><td>Asn</td><td>Arg</td><td>Gly</td><td>Glu</td><td>Cys</td><td></td><td></td><td></td><td></td><td></td><td></td><td></td>

225 230 &lt;210&gt; 68 &lt;2il&gt; 463 &lt;212&gt; PRT &lt;213&gt; Homo sapiens &lt;4068

<td>Underworld 1</td><td>Gili</td><td>Phe</td><td>Gly</td><td>Leu 5</td><td>Cheese</td><td>Trp</td><td>Val</td><td>Phe</td><td>Leu 10</td><td></td><td>Ala</td><td>Leu</td><td>Leu</td><td>Arg 15</td><td>Gly</td>

<td>vai</td><td>Gin</td><td>Cys</td><td>01 .. π twenty</td><td>Val</td><td>Gin</td><td>Leu</td><td>Val</td><td>Glu 25</td><td>Cheese</td><td>Gly</td><td>Gly</td><td>Gly</td><td>Val thirty</td><td>Val</td><td>Glu</td>

PL 214 003 Β1

<td>Pro</td><td>Gly</td><td>Arg 35</td><td>Cheese</td><td>Leu</td><td>Arg</td><td>Leu</td><td>Cheese 40</td><td>Cys</td><td>Thr</td><td>Ala</td><td>Cheese</td><td>Gly 4 5</td><td>Phe</td><td>Thr</td><td>Phe</td>

<td>Cheese</td><td>Cheese</td><td>Tyr</td><td>Gly</td><td>Underworld</td><td>His</td><td>Trp</td><td>Val</td><td>Arg</td><td>Gin</td><td>Ala</td><td>Pro</td><td>Gly</td><td>Lys</td><td>Gly</td><td>Leu</td>

<td></td><td> 50</td><td></td><td></td><td></td><td></td><td> 55</td><td></td><td></td><td></td><td></td><td> 60</td><td></td><td></td><td></td><td></td>

<td>Glu</td><td>Trp</td><td>Val</td><td>Ala</td><td>Val</td><td>How much</td><td>Trp</td><td>Tyr</td><td>Asp</td><td>Gly</td><td>Cheese</td><td>Asn</td><td>Lys</td><td>His</td><td>Tyr</td><td>Ala</td>

<td> 65</td><td></td><td></td><td></td><td></td><td> 70</td><td></td><td></td><td></td><td></td><td> 75</td><td></td><td></td><td></td><td></td><td> 80</td>

<td>Asp</td><td>Cheese</td><td>Ala</td><td>Lys</td><td>Gly</td><td>Arg</td><td>Phe</td><td>Thr</td><td>How much</td><td>Cheese</td><td>Arg</td><td>Asp</td><td>Asn</td><td>Cheese</td><td>Lys</td><td>Asn</td>

<td></td><td></td><td></td><td></td><td> 85</td><td></td><td></td><td></td><td></td><td> 90</td><td></td><td></td><td></td><td></td><td> 95</td><td></td>

<td>Thr</td><td>Leu</td><td>Tyr</td><td>Leu</td><td>Gin</td><td>Underworld</td><td>Asn</td><td>Cheese</td><td>Leu</td><td>Arg</td><td>Ala</td><td>Glu</td><td>Asp</td><td>Thr</td><td>Ala</td><td>Val</td>

<td></td><td></td><td></td><td> 100</td><td></td><td></td><td></td><td></td><td> 105</td><td></td><td></td><td></td><td></td><td> 110</td><td></td><td></td>

<td>Tyr</td><td>Tyr</td><td>Cys</td><td>Ala</td><td>Arg</td><td>Ala</td><td>Gly</td><td>Leu</td><td>Leu</td><td>Gly</td><td>Tyr</td><td>Phe</td><td>Asp</td><td>Tyr</td><td>Trp</td><td>Gly</td>

<td></td><td></td><td> 115</td><td></td><td></td><td></td><td></td><td> 120</td><td></td><td></td><td></td><td></td><td> 125</td><td></td><td></td><td></td>

<td>Gin</td><td>dy</td><td>Thr</td><td>Leu</td><td>Val</td><td>Thr</td><td>Val</td><td>Cheese</td><td>Cheese</td><td>Ala</td><td>Cheese</td><td>Thr</td><td>Bald</td><td>Gly</td><td>Pro</td><td>Cheese</td>

<td></td><td> 130</td><td></td><td></td><td></td><td></td><td> 135</td><td></td><td></td><td></td><td></td><td> 140</td><td></td><td></td><td></td><td></td>

<td>Val</td><td>Phe</td><td>Pro</td><td>Leu</td><td>Ala</td><td>Pro</td><td>Cys</td><td>Cheese</td><td>Arg</td><td>Cheese</td><td>Thr</td><td>Cheese</td><td>Glu</td><td>Cheese</td><td>Thr</td><td>Al. and</td>

<td> 145</td><td></td><td></td><td></td><td></td><td> 150</td><td></td><td></td><td></td><td></td><td> 155</td><td></td><td></td><td></td><td></td><td> 160</td>

<td>Ala</td><td>Leu</td><td>Gly</td><td>Cys</td><td>Leu</td><td>Val</td><td>Lys</td><td>Asp</td><td>Tyr</td><td>Phe</td><td>Pro</td><td>Glu</td><td>Pro</td><td>val</td><td>Thr</td><td>Val</td>

<td></td><td></td><td></td><td></td><td> 165</td><td></td><td></td><td></td><td></td><td> 170</td><td></td><td></td><td></td><td></td><td> 175</td><td></td>

<td>Cheese</td><td>Trp</td><td>Asn</td><td>Cheese</td><td>Gly</td><td>Ala</td><td>Leu</td><td>Thr</td><td>Cheese</td><td>Gly</td><td>Val</td><td>His</td><td>Thr</td><td>Phe</td><td>Pro</td><td>Ala</td>

<td></td><td></td><td></td><td> 180</td><td></td><td></td><td></td><td></td><td> 185</td><td></td><td></td><td></td><td></td><td> 190</td><td></td><td></td>

<td>Val</td><td>Leu</td><td>Gin</td><td>Cheese</td><td>Cheese</td><td>Gly</td><td>Leu</td><td>Tyr</td><td>Cheese</td><td>Leu</td><td>Cheese</td><td>Cheese</td><td>Val</td><td>Val</td><td>Thr</td><td>Val</td>

<td></td><td></td><td> 195</td><td></td><td></td><td></td><td></td><td> 200</td><td></td><td></td><td></td><td></td><td> 205</td><td></td><td></td><td></td>

<td>Pro</td><td>Cheese</td><td>Cheese</td><td>Asn</td><td>Phe</td><td>dy</td><td>Thr</td><td>Gin</td><td>Thr</td><td>Tyr</td><td>Thr</td><td>Cys</td><td>Asn</td><td>Val</td><td>Asp</td><td>His</td>

<td></td><td> 210</td><td></td><td></td><td></td><td></td><td> 215</td><td></td><td></td><td></td><td></td><td> 220</td><td></td><td></td><td></td><td></td>

<td>Lys</td><td>Pro</td><td>Cheese</td><td>Asn</td><td>Thr</td><td>Lys</td><td>Val</td><td>Asp</td><td>Lys</td><td>Thr</td><td>Val</td><td>Glu</td><td>Arg</td><td>Lys</td><td>Cys</td><td>Cys</td>

<td> 225</td><td></td><td></td><td></td><td></td><td> 230</td><td></td><td></td><td></td><td></td><td> 235</td><td></td><td></td><td></td><td></td><td> 240</td>

<td>Val</td><td>Glu</td><td>Cys</td><td>Pro</td><td>Pro</td><td>Cys</td><td>Pro</td><td>Ala</td><td>Pro</td><td>Pro</td><td>val</td><td>Ala</td><td>Gly '</td><td>Pro</td><td>Cheese</td><td>Val</td>

<td></td><td></td><td></td><td></td><td> 245</td><td></td><td></td><td></td><td></td><td> 250</td><td></td><td></td><td></td><td></td><td> 255</td><td></td>

<td>Phe</td><td>Leu</td><td>Phe</td><td>Pro</td><td>Pro</td><td>Lys</td><td>Pro</td><td>Lys</td><td>Asp</td><td>Thr</td><td>Leu</td><td>Underworld</td><td>How much</td><td>Cheese</td><td>Arg</td><td>Thr</td>

<td></td><td></td><td></td><td> 260</td><td></td><td></td><td></td><td></td><td> 265</td><td></td><td></td><td></td><td></td><td> 270</td><td></td><td></td>

<td>Pro</td><td>Glu</td><td>Val</td><td>Thr</td><td>Cys</td><td>Val</td><td>Val</td><td>Val</td><td>Asp</td><td>val</td><td>Cheese</td><td>His</td><td>Glu</td><td>Asp</td><td>Pro</td><td>Glu</td>

<td></td><td></td><td> 275</td><td></td><td></td><td></td><td></td><td> 280</td><td></td><td></td><td></td><td></td><td> 285</td><td></td><td></td><td></td>

<td>val</td><td>Gin</td><td>Phe</td><td>Asn</td><td>Trp</td><td>Tyr</td><td>Val</td><td>Asp</td><td>Gly</td><td>Val</td><td>Glu</td><td>Val</td><td>His</td><td>Asn</td><td>Ala</td><td>Lys</td>

<td></td><td> 290</td><td></td><td></td><td></td><td></td><td> 295</td><td></td><td></td><td></td><td></td><td> 300</td><td></td><td></td><td></td><td></td>

<td>Thr</td><td>Lys</td><td>Pro</td><td>Arg</td><td>Glu</td><td>Glu</td><td>Gin</td><td>Phe</td><td>Asn</td><td>Cheese</td><td>Thr</td><td>Phe</td><td>Arg</td><td>Val</td><td>Val</td><td>Cheese</td>

<td> 305</td><td></td><td></td><td></td><td></td><td> 310</td><td></td><td></td><td></td><td></td><td> 315</td><td></td><td></td><td></td><td></td><td> 320</td>

<td>Val</td><td>Leu</td><td>Thr</td><td>Val</td><td>Val</td><td>His</td><td>Gl.n</td><td>Asp</td><td>Trp</td><td>Leu</td><td>Asn</td><td>Gly</td><td>Lys</td><td>Glu</td><td>Tyr</td><td>Lys</td>

<td></td><td></td><td></td><td></td><td> 325</td><td></td><td></td><td></td><td></td><td> 330</td><td></td><td></td><td></td><td></td><td> 335</td><td></td>

Cys Lys Val Ser Asn Lys Gly Leu Pro Ala Pro Ile Glu Lys Thr Ile

PL 214 003 Β1

<td></td><td></td><td></td><td> 340</td><td></td><td></td><td></td><td></td><td> 345</td><td></td><td></td><td></td><td></td><td> 350</td><td></td><td></td>

<td>Cheese</td><td>Lys</td><td>Thr</td><td>Lys</td><td>Gly</td><td>G1 n</td><td>Pro</td><td>Arg</td><td>Glu</td><td>Pro</td><td>Gin</td><td>Val</td><td>Tyr</td><td>Thr</td><td>Leu</td><td>Pro</td>

<td></td><td></td><td> 355</td><td></td><td></td><td></td><td></td><td> 360</td><td></td><td></td><td></td><td></td><td> 365</td><td></td><td></td><td></td>

<td>Pro</td><td>Cheese</td><td>Arg</td><td>Glu</td><td>Glu</td><td>Underworld</td><td>Thr</td><td>Lys</td><td>Asn</td><td>Gin</td><td>Val</td><td>Cheese</td><td>Leu</td><td>Thr</td><td>Cys</td><td>Leu</td>

<td></td><td> 370</td><td></td><td></td><td></td><td></td><td> 375</td><td></td><td></td><td></td><td></td><td> 380</td><td></td><td></td><td></td><td></td>

<td>Val</td><td>Lys</td><td>Gly</td><td>Phe</td><td>Tyr</td><td>Pro</td><td>Cheese</td><td>Asp</td><td>How much</td><td>Ala</td><td>Val</td><td>Glu</td><td>Trp</td><td>Glu</td><td>Cheese</td><td>Asn</td>

<td> 385</td><td></td><td></td><td></td><td></td><td> 390</td><td></td><td></td><td></td><td></td><td> 395</td><td></td><td></td><td></td><td></td><td> 400</td>

<td>dy</td><td>Gin</td><td>Pro</td><td>Glu</td><td>Asn</td><td>Asn</td><td>Tyr</td><td>Lys</td><td>Thr</td><td>Thr</td><td>Pro</td><td>Pro</td><td>Underworld</td><td>Leu</td><td>Asp</td><td>Cheese</td>

<td></td><td></td><td></td><td></td><td> 405</td><td></td><td></td><td></td><td></td><td> 410</td><td></td><td></td><td></td><td></td><td> 415</td><td></td>

<td>Asp</td><td>G1 y</td><td>Cheese</td><td>Phe</td><td>Phe</td><td>Leu</td><td>Tyr</td><td>Cheese</td><td>Lys</td><td>Leu</td><td>Thr</td><td> 1.</td><td>Asp</td><td>Lys</td><td>Cheese</td><td>Arg</td>

<td></td><td></td><td></td><td> 420</td><td></td><td></td><td></td><td></td><td> 425</td><td></td><td></td><td></td><td></td><td> 430</td><td></td><td></td>

<td>Trp</td><td>Gin</td><td>Gin</td><td>Gly</td><td>Asn</td><td>Val</td><td>Phe</td><td>Cheese</td><td>Cys</td><td>Cheese</td><td>Val</td><td>Underworld</td><td>His</td><td>Glu</td><td>Al a</td><td>Leu</td>

<td></td><td></td><td> 435</td><td></td><td></td><td></td><td></td><td> 440</td><td></td><td></td><td></td><td></td><td> 445</td><td></td><td></td><td></td>

<td>His</td><td>Asn</td><td>His</td><td>Tyr</td><td>Thr</td><td>Gin</td><td>Lys</td><td>Cheese</td><td>Leu</td><td>Cheese</td><td>Leu</td><td>Cheese</td><td>Pro</td><td>Gly</td><td>Lys</td><td></td>

450 455 460 &lt;210&gt; 69 &lt;211&gt; 234 &lt;212&gt; PRT &lt;213&gt; Homo sapiens &lt;400&gt; 69

<td>Underworld 1</td><td>Glu</td><td>Thr</td><td>Pro</td><td>Ala 5</td><td>Gin</td><td>Leu</td><td>Leu</td><td>Phe</td><td>Leu 10</td><td>Leu</td><td>Leu</td><td>Leu</td><td>Trp</td><td>Leu 15</td><td>Pro</td>

<td>Asp</td><td>Thr</td><td>Thr</td><td>Gly</td><td>Glu</td><td>How much</td><td>Val</td><td>Leu</td><td>Thr</td><td>Gin</td><td>Cheese</td><td>Pro</td><td>Gly</td><td>Thr</td><td>Leu</td><td>Cheese</td>

<td></td><td></td><td></td><td> 20</td><td></td><td></td><td></td><td></td><td> 25</td><td></td><td></td><td></td><td></td><td> 30</td><td></td><td></td>

<td>Leu</td><td>Cheese</td><td>Pro</td><td>Gly</td><td>Glu</td><td>Arg</td><td>Ala</td><td>Thr</td><td>Leu</td><td>Cheese</td><td>Cys</td><td>Arg</td><td>Ala</td><td>Cheese</td><td>Gin</td><td>Cheese</td>

<td></td><td></td><td> 35</td><td></td><td></td><td></td><td></td><td> 40</td><td></td><td></td><td></td><td></td><td> 45</td><td></td><td></td><td></td>

<td>Val</td><td>Cheese</td><td>Cheese</td><td>Tyr</td><td>Leu</td><td>Ala</td><td>Trp</td><td>Tyr</td><td>Gin</td><td>Gin</td><td>Lys</td><td>Pro</td><td>Gly</td><td>Gin</td><td>Ala</td><td>Pro</td>

<td></td><td> 50</td><td></td><td></td><td></td><td></td><td> 55</td><td></td><td></td><td></td><td></td><td> 60</td><td></td><td></td><td></td><td></td>

<td>Arg</td><td>Pro</td><td>Leu</td><td>How much</td><td>Tyr</td><td>Gly</td><td>Val</td><td>Cheese</td><td>Cheese</td><td>Arg</td><td>Ala</td><td>Thr</td><td>Gly</td><td>How much</td><td>Pro</td><td>Asp</td>

<td> 65</td><td></td><td></td><td></td><td></td><td> 70</td><td></td><td></td><td></td><td></td><td> 75</td><td></td><td></td><td></td><td></td><td> 80</td>

<td>Arg</td><td>Phe</td><td>Cheese</td><td>Gly</td><td>Cheese</td><td>Gly</td><td>Cheese</td><td>Gly</td><td>Thr</td><td>Asp</td><td>Phe</td><td>Thr</td><td>Leu</td><td>Thr</td><td>How much</td><td>Cheese</td>

<td></td><td></td><td></td><td></td><td> 85</td><td></td><td></td><td></td><td></td><td> 90</td><td></td><td></td><td></td><td></td><td> 95</td><td></td>

<td>Arg</td><td>Leu</td><td>Glu</td><td>Pro</td><td>Glu</td><td>Asp</td><td>Phe</td><td>Ala</td><td>Val</td><td>Tyr</td><td>Tyr</td><td>Cys</td><td>Gin</td><td>Gin</td><td>Tyr</td><td>Gly</td>

<td></td><td></td><td></td><td> 100</td><td></td><td></td><td></td><td></td><td> 105</td><td></td><td></td><td></td><td></td><td> 110</td><td></td><td></td>

<td>How much</td><td>Cheese</td><td>Pro</td><td>Phe</td><td>Thr</td><td>Phe</td><td>Gly</td><td>Pro</td><td>Gly</td><td>Thr</td><td>Lys</td><td>Val</td><td>Asp</td><td>How much</td><td>Lys</td><td>Arg</td>

<td></td><td></td><td> 115</td><td></td><td></td><td></td><td></td><td> 120</td><td></td><td></td><td></td><td></td><td> 125</td><td></td><td></td><td></td>

<td>Thr</td><td>Val</td><td>Ala</td><td>Ala</td><td>Pro</td><td>Cheese</td><td>Val</td><td>Phe</td><td>How much</td><td>Phe</td><td>Pro</td><td>Pro</td><td>Cheese</td><td>Asp</td><td>Glu</td><td>Gin</td>

<td></td><td> 130</td><td></td><td></td><td></td><td></td><td> 135</td><td></td><td></td><td></td><td></td><td> 140</td><td></td><td></td><td></td><td></td>

Leu Lys Ser Gly Thr Ala Ser Val Val Cys Leu Leu Asn Asn Phe Tyr

PL 214 003 Β1

<td> 145</td><td></td><td></td><td></td><td></td><td> 150</td><td></td><td></td><td></td><td></td><td> 155</td><td></td><td></td><td></td><td></td><td> 160</td>

<td>Pro</td><td>Arg</td><td>Glu</td><td>Ala</td><td>Lys</td><td>Val</td><td>Gin</td><td>Trp</td><td>Lys</td><td>Val</td><td>Asp</td><td>Asn</td><td>Ar a</td><td>Leu</td><td>Gin</td><td>Cheese</td>

<td></td><td></td><td></td><td></td><td> 165</td><td></td><td></td><td></td><td></td><td> 170</td><td></td><td></td><td></td><td></td><td> 175</td><td></td>

<td>Gly</td><td>Asn</td><td>Cheese</td><td>Gin</td><td>Glu</td><td>Cheese</td><td>Val</td><td>Thr</td><td>Glu</td><td>Gin</td><td>Asp</td><td>Cheese</td><td>Lys</td><td>Asp</td><td>Cheese</td><td>Thr</td>

<td></td><td></td><td></td><td> 180</td><td></td><td></td><td></td><td></td><td> 185</td><td></td><td></td><td></td><td></td><td> 190</td><td></td><td></td>

<td>Tyr</td><td>Cheese</td><td>Leu</td><td>Cheese</td><td>Cheese</td><td>Thr</td><td>Leu</td><td>Thr</td><td>Leu</td><td>Cheese</td><td>Lys</td><td>Ala</td><td>Asp</td><td>Tyr</td><td>Glu</td><td>Lys</td>

<td></td><td></td><td> 195</td><td></td><td></td><td></td><td></td><td> 200</td><td></td><td></td><td></td><td></td><td> 205</td><td></td><td></td><td></td>

<td>His</td><td>Lys</td><td>Val</td><td>Tyr</td><td>Ala</td><td>Cys</td><td>Glu</td><td>Val</td><td>Thr</td><td>His</td><td>Gin</td><td>Gly</td><td>Leu</td><td>Cheese</td><td>Ss r</td><td>Pro</td>

<td></td><td> 210</td><td></td><td></td><td></td><td></td><td> 215</td><td></td><td></td><td></td><td></td><td> 220</td><td></td><td></td><td></td><td></td>

<td>Val</td><td>Thr</td><td>Lys</td><td>Cheese</td><td>Phe</td><td>Asn</td><td>Arg</td><td>Gly</td><td>Glu</td><td>cys</td><td></td><td></td><td></td><td></td><td></td><td></td>

225 230 &lt;210&gt; 70 &lt;211&gt; 451 &lt;212&gt; PRT &lt;213&gt; Homo sapiens &lt;400&gt; 70

<td>Gin 1</td><td>Val</td><td>Gin</td><td>Leu</td><td>Val 5</td><td>Glu</td><td>Cheese</td><td>Gly</td><td colspan="3">Gly Gly Val 10</td><td>Val</td><td>Gin</td><td>Pro</td><td>Gly 15</td><td>Arg</td>

<td>Cheese</td><td>Leu</td><td>Arg</td><td>Leu</td><td>Cheese</td><td>Cys</td><td>Ala</td><td>Ala</td><td>Cheese</td><td>Gly</td><td>Phe</td><td>Thr</td><td>Phe</td><td>Cheese</td><td>Cheese</td><td>Tyr</td>

<td></td><td></td><td></td><td> 20</td><td></td><td></td><td></td><td></td><td> 25</td><td></td><td></td><td></td><td></td><td> 30</td><td></td><td></td>

<td>Gly</td><td>Underworld</td><td>His</td><td>Trp</td><td>Val</td><td>Arg</td><td>Gin</td><td>Ala</td><td>Pro</td><td>Gly</td><td>Lys</td><td>Gly</td><td>Leu</td><td>Glu</td><td>Trp</td><td>Val</td>

<td></td><td></td><td> 35</td><td></td><td></td><td></td><td></td><td> 40</td><td></td><td></td><td></td><td></td><td> 4 5</td><td></td><td></td><td></td>

<td>Ala</td><td>Val</td><td>How much</td><td>Trp</td><td>Tyr</td><td>Asp</td><td>Gly</td><td>Cheese</td><td>Asn</td><td>Lys</td><td>Tyr</td><td>Tyr</td><td>Al a</td><td>Asp</td><td>Cheese</td><td>Val</td>

<td></td><td> 50</td><td></td><td></td><td></td><td></td><td> 55</td><td></td><td></td><td></td><td></td><td> 60</td><td></td><td></td><td></td><td></td>

<td>Lys</td><td>Gly</td><td>Arg</td><td>Phe</td><td>Thr</td><td>How much</td><td>Cheese</td><td>Arg</td><td>Asp</td><td>Asn</td><td>Cheese</td><td>Lys</td><td>Asn</td><td>Thr</td><td>Leu</td><td>Tyr</td>

<td> 65</td><td></td><td></td><td></td><td></td><td> 70</td><td></td><td></td><td></td><td></td><td> 75</td><td></td><td></td><td></td><td></td><td> 80</td>

<td>Leu</td><td>Gin</td><td>Underworld</td><td>Asn</td><td>Cheese</td><td>Leu</td><td>Arg</td><td>Ala</td><td>Glu</td><td>Asp</td><td>Thr</td><td>Ala</td><td>Val</td><td>Tyr</td><td>Tyr</td><td>cys</td>

<td></td><td></td><td></td><td></td><td> 85</td><td></td><td></td><td></td><td></td><td> 90</td><td></td><td></td><td></td><td></td><td> 95</td><td></td>

<td>Ala</td><td>Arg</td><td>Asp</td><td>Pro</td><td>Arg</td><td>Gly</td><td>Ala</td><td>Thr</td><td>Leu</td><td>Tyr</td><td>Tyr</td><td>Tyr</td><td>Tyr</td><td>Tyr</td><td>Gly</td><td>Underworld</td>

<td></td><td></td><td></td><td> 100</td><td></td><td></td><td></td><td></td><td> 105</td><td></td><td></td><td></td><td></td><td> 110</td><td></td><td></td>

<td>Asp</td><td>Val</td><td>Trp</td><td>Gly</td><td>Gin</td><td>Gly</td><td>Thr</td><td>Thr</td><td>Val</td><td>Thr</td><td>Val</td><td>Cheese</td><td>Cheese</td><td>Ala</td><td>Cheese</td><td>Thr</td>

<td></td><td></td><td> 115</td><td></td><td></td><td></td><td></td><td> 120</td><td></td><td></td><td></td><td></td><td> 125</td><td></td><td></td><td></td>

<td>Lys</td><td>Gly</td><td>Pro</td><td>Cheese</td><td>Val</td><td>Phe</td><td>Pro</td><td>Leu</td><td>Ala</td><td>Pro</td><td>Cys</td><td>Cheese</td><td>Arg</td><td>Cheese</td><td>Thr</td><td>Cheese</td>

<td></td><td> 130</td><td></td><td></td><td></td><td></td><td> 135</td><td></td><td></td><td></td><td></td><td> 140</td><td></td><td></td><td></td><td></td>

<td>Glu</td><td>Cheese</td><td>Thr</td><td>Ala</td><td>Ala</td><td>Leu</td><td>Gly</td><td>Cys</td><td>Leu</td><td>Val</td><td>Lys</td><td>Asp</td><td>Tyr</td><td>Phe</td><td>Pro</td><td>Glu</td>

<td> 145</td><td></td><td></td><td></td><td></td><td> 150</td><td></td><td></td><td></td><td></td><td> 15 5</td><td></td><td></td><td></td><td></td><td> 160</td>

<td>Pro</td><td>Val</td><td>Thr</td><td>Val</td><td>Cheese</td><td>Trp</td><td>Asn</td><td>Cheese</td><td>dy</td><td>Ala</td><td>Leu</td><td>Thr</td><td>Cheese</td><td>Gly</td><td>Vai</td><td>His</td>

<td></td><td></td><td></td><td></td><td> 165</td><td></td><td></td><td></td><td></td><td> 170</td><td></td><td></td><td></td><td></td><td> 175</td><td></td>

Thr Phe Pro Ala Val Leu Gin Ser Ser Gly Leu Tyr Ser Leu Ser Ser

PL 214 003 Β1

<td></td><td colspan="5"> 180</td><td colspan="7"> 185</td><td colspan="3"> 190</td>

<td>Val</td><td>Val</td><td>Thr</td><td>Val</td><td>Pro</td><td>Cheese</td><td>Cheese</td><td>Asn</td><td>Phe</td><td>Gly</td><td>Thr</td><td>Gin</td><td>Thr</td><td>Tyr</td><td>Thr</td><td>Cys</td>

<td></td><td></td><td> 195</td><td></td><td></td><td></td><td></td><td> 200</td><td></td><td></td><td></td><td></td><td> 205</td><td></td><td></td><td></td>

<td>Asn</td><td>Val</td><td>Asp</td><td>His</td><td>Lys</td><td>Pro</td><td>Cheese</td><td>Asn</td><td>Thr</td><td>Lys</td><td>Val</td><td>Asp</td><td>Lys</td><td>Thr</td><td>val</td><td>Glu</td>

<td></td><td> 210</td><td></td><td></td><td></td><td></td><td> 215</td><td></td><td></td><td></td><td></td><td> 220</td><td></td><td></td><td></td><td></td>

<td>Arg</td><td>Lys</td><td>Cys</td><td>Cys</td><td>Val</td><td>Glu</td><td>Cys</td><td>Pro</td><td>Pro</td><td>Cys</td><td>Pro</td><td>Ala</td><td>Pro</td><td>Pro</td><td>Val</td><td>Ala</td>

<td> 225</td><td></td><td></td><td></td><td></td><td> 230</td><td></td><td></td><td></td><td></td><td> 235</td><td></td><td></td><td></td><td></td><td> 240</td>

<td>Gly</td><td>Pro</td><td>Cheese</td><td>Val</td><td>Phe</td><td>Leu</td><td>Phe</td><td>Pro</td><td>Pro</td><td>Lys</td><td>Pro</td><td>Lys</td><td>Asp</td><td>Thr</td><td>Leu</td><td>Underworld</td>

<td></td><td></td><td></td><td></td><td> 245</td><td></td><td></td><td></td><td></td><td> 250</td><td></td><td></td><td></td><td></td><td> 255</td><td></td>

<td>He</td><td>Cheese</td><td>Arg</td><td>Thr</td><td>Pro</td><td>Glu</td><td>Val</td><td>Thr</td><td>cys</td><td>Val</td><td>Val</td><td>Val</td><td>Asp</td><td>Val</td><td>Cheese</td><td>His</td>

<td></td><td></td><td></td><td> 260</td><td></td><td></td><td></td><td></td><td> 265</td><td></td><td></td><td></td><td></td><td> 270</td><td></td><td></td>

<td>Glu</td><td>Asp</td><td>Pro</td><td>Glu</td><td>Val</td><td>Gin</td><td>Phe</td><td>Asn</td><td>Trp</td><td>Tyr</td><td>Val</td><td>Asp</td><td>Gly</td><td>Val</td><td>Glu</td><td>Val</td>

<td></td><td></td><td> 275</td><td></td><td></td><td></td><td></td><td> 280</td><td></td><td></td><td></td><td></td><td> 285</td><td></td><td></td><td></td>

<td>His</td><td>Asn</td><td>Al a</td><td>Lys</td><td>Thr</td><td>Lys</td><td>Pro</td><td>Arg</td><td>Glu</td><td>Glu</td><td>Gin</td><td>Phe</td><td>Asn</td><td>Cheese</td><td>Thr</td><td>Phe</td>

<td></td><td> 290</td><td></td><td></td><td></td><td></td><td> 295</td><td></td><td></td><td></td><td></td><td> 300</td><td></td><td></td><td></td><td></td>

<td>Arg</td><td>Val</td><td>Val</td><td>Cheese</td><td>Vai</td><td>Leu</td><td>Thr</td><td>Val</td><td>Val</td><td>His</td><td>Gin</td><td>Asp</td><td>Trp</td><td>Leu</td><td>Asn</td><td>Gly</td>

<td> 305</td><td></td><td></td><td></td><td></td><td> 310</td><td></td><td></td><td></td><td></td><td> 315</td><td></td><td></td><td></td><td></td><td> 3 20</td>

<td>Lys</td><td>Glu</td><td>Tyr</td><td>Lys</td><td>Cys</td><td>Lys</td><td>Val</td><td>Cheese</td><td>Asn</td><td>Lys</td><td>Gly</td><td>Leu</td><td>Pro</td><td>Ala</td><td>Pro</td><td>How much</td>

<td></td><td></td><td></td><td></td><td> 325</td><td></td><td></td><td></td><td></td><td> 330</td><td></td><td></td><td></td><td></td><td> 335</td><td></td>

<td>Glu</td><td>Lys</td><td>Thr</td><td>How much</td><td>Cheese</td><td>Lys</td><td>Thr</td><td>Lys</td><td>Gly</td><td>Gin</td><td>Pro</td><td>Arg</td><td>Glu</td><td>Pro</td><td>Gin</td><td>Val</td>

<td></td><td></td><td></td><td> 340</td><td></td><td></td><td></td><td></td><td> 345</td><td></td><td></td><td></td><td></td><td> 350</td><td></td><td></td>

<td>Tyr</td><td>Thr</td><td>Leu</td><td>Pro</td><td>Pro</td><td>Cheese</td><td>Arg</td><td>Glu</td><td>Glu</td><td>Underworld</td><td>Thr</td><td>Lys</td><td>Asn</td><td>Gin</td><td>Val</td><td>Cheese</td>

<td></td><td></td><td> 355</td><td></td><td></td><td></td><td></td><td> 360</td><td></td><td></td><td></td><td></td><td> 365</td><td></td><td></td><td></td>

<td>Leu</td><td>Thr</td><td>cys</td><td>Leu</td><td>Va 1</td><td>Lys</td><td>Gly</td><td>Phe</td><td>Tyr</td><td>Pro</td><td>Cheese</td><td>Asp</td><td>How much</td><td>Ala</td><td>Val</td><td>Glu</td>

<td></td><td> 370</td><td></td><td></td><td></td><td></td><td> 375</td><td></td><td></td><td></td><td></td><td> 380</td><td></td><td></td><td></td><td></td>

<td>Trp</td><td>Glu</td><td>Cheese</td><td>Asn</td><td>Gly</td><td>Gin</td><td>Pro</td><td>Glu</td><td>Asn</td><td>Asn</td><td>Tyr</td><td>Lys</td><td>Thr</td><td>Thr</td><td>Pro</td><td>Pro</td>

<td> 385</td><td></td><td></td><td></td><td></td><td> 390</td><td></td><td></td><td></td><td></td><td> 395</td><td></td><td></td><td></td><td></td><td> 400</td>

<td>Underworld</td><td>Leu</td><td>Asp</td><td>Cheese</td><td>Asp</td><td>Gly</td><td>Cheese</td><td>Phe</td><td>Phe</td><td>Leu</td><td>Tyr</td><td>Cheese</td><td>Lys</td><td>Leu</td><td>Thr</td><td>vai</td>

<td></td><td></td><td></td><td></td><td> 405</td><td></td><td></td><td></td><td></td><td> 410</td><td></td><td></td><td></td><td></td><td> 415</td><td></td>

<td>Asp</td><td>Lys</td><td>Cheese</td><td>Arg</td><td>Trp</td><td>Gin</td><td>Gin</td><td>Giy</td><td>Asn</td><td>Val</td><td>Phe</td><td>Cheese</td><td>Cys</td><td>Cheese</td><td>Val</td><td>Underworld</td>

<td></td><td></td><td></td><td> 420</td><td></td><td></td><td></td><td></td><td> 425</td><td></td><td></td><td></td><td></td><td> 430</td><td></td><td></td>

<td>His</td><td>Glu</td><td>Ala</td><td>Leu</td><td>His</td><td>Asn</td><td>His</td><td>Tyr</td><td>Thr</td><td>Gin</td><td>Lys</td><td>Cheese</td><td>Leu</td><td>Cheese</td><td>Leu</td><td>Cheese</td>

<td></td><td></td><td> 435</td><td></td><td></td><td></td><td></td><td> 440</td><td></td><td></td><td></td><td></td><td> 445</td><td></td><td></td><td></td>

Pro Gly Lys

450

<td> &lt;210&gt;</td><td> 71</td>

<td> &lt;211&gt;</td><td> 214</td>

<td> &lt;212&gt;</td><td>PRT</td>

<td> &lt;213&gt;</td><td>Homo</td>

sapiens

PL 214 003 Β1 &lt;400 71

<td>Asp 1</td><td>How much</td><td>Gin</td><td>Underworld</td><td>Thr 5</td><td>Gin.</td><td>Cheese</td><td>Pro</td><td>Cheese</td><td>Cheese 10</td><td>Leu</td><td>Cheese</td><td>Ala</td><td>Cheese</td><td>Val 15</td><td>Gly</td>

<td>Asp</td><td>Arg</td><td>Val</td><td>Thr</td><td>How much</td><td>Thr</td><td>Cys</td><td>Arg</td><td>Ala</td><td>Cheese</td><td>Gin</td><td>Cheese</td><td>How much</td><td>Asn</td><td>Cheese</td><td>Tyr</td>

<td></td><td></td><td></td><td> 20</td><td></td><td></td><td></td><td></td><td> 25</td><td></td><td></td><td></td><td></td><td> 30</td><td></td><td></td>

<td>Leu</td><td>Asp</td><td>Trp</td><td>Tyr</td><td>Gin</td><td>Gin</td><td>Lys</td><td>Pro</td><td>Gly</td><td>Lys</td><td>Ala</td><td>Pro</td><td>Lys</td><td>Leu</td><td>Leu</td><td>1 le</td>

<td></td><td></td><td> 35</td><td></td><td></td><td></td><td></td><td> 40</td><td></td><td></td><td></td><td></td><td> 45</td><td></td><td></td><td></td>

<td>Tyr</td><td> 7^.1.</td><td>Ala</td><td>Cheese</td><td>Cheese</td><td>Leu</td><td>Gin</td><td>Cheese</td><td>Gly</td><td>Val</td><td>Pro</td><td>Cheese</td><td>Arg</td><td>Phe</td><td>Cheese</td><td>Gly</td>

<td></td><td> 50</td><td></td><td></td><td></td><td></td><td> 55</td><td></td><td></td><td></td><td></td><td> 60</td><td></td><td></td><td></td><td></td>

<td>Cheese</td><td>Giy</td><td>Cheese</td><td>Gly</td><td>Thr</td><td>Asp</td><td>Phe</td><td>Thr</td><td>Leu</td><td>Thr</td><td>how much</td><td>Cheese</td><td>Cheese</td><td>Leu</td><td>Gin</td><td>Pro</td>

<td> 65</td><td></td><td></td><td></td><td></td><td> 70</td><td></td><td></td><td></td><td></td><td> 75</td><td></td><td></td><td></td><td></td><td> 80</td>

<td>Glu</td><td>Asp</td><td>Phe</td><td>Ala</td><td>Thr</td><td>Tyr</td><td>Tyr</td><td>cys</td><td>Gin</td><td>Gin</td><td>Tyr</td><td>Tyr</td><td>Cheese</td><td>Thr</td><td>Pro</td><td>Phe</td>

<td></td><td></td><td></td><td></td><td> 85</td><td></td><td></td><td></td><td></td><td> 90</td><td></td><td></td><td></td><td></td><td> 95</td><td></td>

<td>Thr</td><td>Phe</td><td>Gly</td><td>Pro</td><td>Gly</td><td>Thr</td><td>Lys</td><td>Val</td><td>Glu</td><td>How much</td><td>Lys</td><td>Arg</td><td>Thr</td><td>Val</td><td>Al a</td><td>Ala</td>

<td></td><td></td><td></td><td> 100</td><td></td><td></td><td></td><td></td><td> 105</td><td></td><td></td><td></td><td></td><td> 110</td><td></td><td></td>

<td>Pro</td><td>Cheese</td><td>Val</td><td>Phe</td><td>How much</td><td>Phe</td><td>Pro</td><td>Pro</td><td>Cheese</td><td>Asp</td><td>Glu</td><td>Gin</td><td>Leu</td><td>Lys</td><td>Cheese</td><td>Gly</td>

<td></td><td></td><td> 115</td><td></td><td></td><td></td><td></td><td> 120</td><td></td><td></td><td></td><td></td><td> 125</td><td></td><td></td><td></td>

<td>Thr</td><td>Al a</td><td>Cheese</td><td>Val</td><td>Val</td><td>Cys</td><td>Leu</td><td>Leu</td><td>Asn</td><td>Asn</td><td>Phe</td><td>Tyr</td><td>Pro</td><td>Arg</td><td>Glu</td><td>Ala</td>

<td></td><td> 130</td><td></td><td></td><td></td><td></td><td> 135</td><td></td><td></td><td></td><td></td><td> '14 0</td><td></td><td></td><td></td><td></td>

<td>Lys</td><td>Val</td><td>Gin</td><td>Trp</td><td>Lys</td><td>Val</td><td>Asp</td><td>Asn</td><td>Ala</td><td>Leu</td><td>Gin</td><td>Cheese</td><td>Gly</td><td>Asn</td><td>Cheese</td><td>Gin</td>

<td> 145</td><td></td><td></td><td></td><td></td><td> 150</td><td></td><td></td><td></td><td></td><td> 155</td><td></td><td></td><td></td><td></td><td> 160</td>

<td>Glu</td><td>Cheese</td><td>Val</td><td>Thr</td><td>Glu</td><td>Gin</td><td>Asp</td><td>Cheese</td><td>Lys</td><td>Asp</td><td>Cheese</td><td>Thr</td><td>Tyr</td><td>Cheese</td><td>Leu</td><td>Cheese</td>

<td></td><td></td><td></td><td></td><td> 165</td><td></td><td></td><td></td><td></td><td> 170</td><td></td><td></td><td></td><td></td><td> 175</td><td></td>

<td>Cheese</td><td>Thr</td><td>Leu</td><td>Thr</td><td>Leu</td><td>Cheese</td><td>Lys</td><td>Ala</td><td>Asp</td><td>Tyr</td><td>Glu</td><td>Lys</td><td>His</td><td>Lys</td><td>Val</td><td>Tyr</td>

<td></td><td></td><td></td><td> 180</td><td></td><td></td><td></td><td></td><td> 185</td><td></td><td></td><td></td><td></td><td> 190</td><td></td><td></td>

<td>Ala</td><td>Cys</td><td>Glu</td><td>Val</td><td>Thr</td><td>His</td><td>Gin</td><td>Gly</td><td>Leu</td><td>Cheese</td><td>Cheese</td><td>Pro</td><td>Val</td><td>Thr</td><td>Lys</td><td>Cheese</td>

<td></td><td></td><td> 195</td><td></td><td></td><td></td><td></td><td> 200</td><td></td><td></td><td></td><td></td><td> 205</td><td></td><td></td><td></td>

<td>Phe</td><td>Asn</td><td>Arg</td><td>Gly</td><td>Glu</td><td>Cys</td><td></td><td></td><td></td><td></td><td></td><td></td><td></td><td></td><td></td><td></td>

210 &lt;210&gt; 72 &lt;211&gt; 89 &lt;212&gt; PRT &lt;213&gt; Homo sapiens &lt;400&gt; 72

<td>Gly 1</td><td colspan="2">Val Val</td><td>Gin</td><td>Pro 5</td><td>Gly</td><td>Arg</td><td>Cheese</td><td colspan="3">Leu Arg Leu 10</td><td>Cheese</td><td>Cys</td><td>Ala</td><td>A.1 3 15</td><td>Cheese</td>

<td>Gly</td><td>Phe</td><td>Thr</td><td>Phe</td><td>Cheese</td><td>Cheese</td><td>Tyr</td><td>Gly</td><td>M m 4 ”iti U</td><td>His</td><td>Trp</td><td>vai</td><td>Arg</td><td>Gin</td><td></td><td>Pro</td>

<td></td><td></td><td></td><td> 20</td><td></td><td></td><td></td><td></td><td> 25</td><td></td><td></td><td></td><td></td><td> 30</td><td></td><td></td>

Gly Lys Gly Leu Glu Trp Val Ala Val Ile Trp Tyr Asp Gly Ser Asn

PL 214 003 Β1

<td></td><td></td><td> 35</td><td></td><td></td><td></td><td></td><td> 40</td><td></td><td></td><td></td><td></td><td> 45</td><td></td><td></td><td></td>

<td>Lys</td><td>Tyr 50</td><td>Tyr</td><td>Ala</td><td>Asp</td><td>Cheese</td><td>Val 55</td><td>Lys</td><td>Gly</td><td>Arg</td><td>Phe</td><td>Thr 60</td><td>How much</td><td>Cheese</td><td>Arg</td><td>Asp</td>

<td>Asn 65</td><td>Cheese</td><td>Lys</td><td>Asn</td><td>Thr</td><td>Leu 70</td><td>Tyr</td><td>Leu</td><td>Gin</td><td>Underworld</td><td>Asn 75</td><td>Cheese</td><td>Leu</td><td>Arg</td><td>Alley</td><td>Glu 80</td>

<td>Asp</td><td>Thr</td><td>Ala</td><td>Val</td><td>Tyr</td><td>Tyr</td><td>Cys</td><td>Ala</td><td>Arg</td><td></td><td></td><td></td><td></td><td></td><td></td><td></td>

&lt;210&gt; 73 &lt;211&gt; 169 &lt;212&gt; PRT &lt;213&gt; Homo sapiens &lt;400&gt; 73

<td colspan="2">Gly Val</td><td>Val</td><td>Gin</td><td>Pro 5</td><td colspan="2">Gly Arg</td><td colspan="2">Leu cheese</td><td>Arg 10</td><td>Leu</td><td>Cheese</td><td>Cys</td><td>Ala</td><td>Ala 15</td><td>Cheese</td>

<td>Gly</td><td>Phe</td><td>Thr</td><td>Phe</td><td>Cheese</td><td>Cheese</td><td>Tyr</td><td>Gly</td><td>Underworld</td><td>His</td><td>Trp</td><td>val</td><td>Arg</td><td>Gin</td><td>Ala</td><td>Pro</td>

<td></td><td></td><td></td><td> 20</td><td></td><td></td><td></td><td></td><td> 25</td><td></td><td></td><td></td><td></td><td> 30</td><td></td><td></td>

<td>Gly</td><td>Lys</td><td>Gly</td><td>Leu</td><td>Glu</td><td>Trp</td><td>Val</td><td>Ala</td><td>Val</td><td>How much</td><td>Trp</td><td>Tyr</td><td>Asp</td><td>Gly</td><td>Cheese</td><td>Asn</td>

<td></td><td></td><td> 35</td><td></td><td></td><td></td><td></td><td> 40</td><td></td><td></td><td></td><td></td><td> 45</td><td></td><td></td><td></td>

<td>Lys</td><td>Tyr</td><td>Tyr</td><td>Ala</td><td>Asp</td><td>Cheese</td><td>Val</td><td>Lys</td><td>Gly</td><td>Arg</td><td>Phe</td><td>Thr</td><td>How much</td><td>Cheese</td><td>Arg</td><td>Asp</td>

<td></td><td> 50</td><td></td><td></td><td></td><td></td><td> 55</td><td></td><td></td><td></td><td></td><td> 60</td><td></td><td></td><td></td><td></td>

<td>Asn</td><td>Cheese</td><td>Lys</td><td>Asn</td><td>Thr</td><td>Leu</td><td>Tyr</td><td>Leu</td><td>Gin</td><td>Underworld</td><td>Asn</td><td>Cheese</td><td>Leu</td><td>Arg</td><td>Ala</td><td>Glu</td>

<td> 65</td><td></td><td></td><td></td><td></td><td> 70</td><td></td><td></td><td></td><td></td><td> 75</td><td></td><td></td><td></td><td></td><td> 80</td>

<td>Asp</td><td>Thr</td><td>Ala</td><td>Val</td><td>Tyr</td><td>Tyr</td><td>Cys</td><td>Ala</td><td>Arg</td><td>Gly</td><td>Ala</td><td>Arg</td><td>How much</td><td>Background</td><td>Thr</td><td>Pro</td>

<td></td><td></td><td></td><td></td><td> 85</td><td></td><td></td><td></td><td></td><td> 90</td><td></td><td></td><td></td><td></td><td> 95</td><td></td>

<td>Cys</td><td>Underworld</td><td>Asp</td><td>Val</td><td>Trp</td><td>Gly</td><td>Gin</td><td>Gly</td><td>Thr</td><td>Thr</td><td>val</td><td>Thr</td><td>val</td><td>Cheese</td><td>Cheese</td><td>Ala</td>

<td></td><td></td><td></td><td> 100</td><td></td><td></td><td></td><td></td><td> 105</td><td></td><td></td><td></td><td></td><td> 110</td><td></td><td></td>

<td>Cheese</td><td>Thr</td><td>Lys</td><td>Gly</td><td>Pro</td><td>Cheese</td><td>Val</td><td>Phe</td><td>Pro</td><td>Leu</td><td>Ala</td><td>Pro</td><td>Cys</td><td>Cheese</td><td>Arg</td><td>Cheese</td>

<td></td><td></td><td> 115</td><td></td><td></td><td></td><td></td><td> 120</td><td></td><td></td><td></td><td></td><td> 125</td><td></td><td></td><td></td>

<td>Thr</td><td>Cheese</td><td>Glu</td><td>Cheese</td><td>Thr</td><td>Ala</td><td>Ala</td><td>Leu</td><td>Gly</td><td>Cys</td><td>Leu</td><td>Val</td><td>Lys</td><td>Asp</td><td>Tyr</td><td>Phe</td>

<td></td><td> 130</td><td></td><td></td><td></td><td></td><td> 135</td><td></td><td></td><td></td><td></td><td> 140</td><td></td><td></td><td></td><td></td>

<td>Pro</td><td>Glu</td><td>Pro</td><td>Val</td><td>Thr</td><td>Val</td><td>Cheese</td><td>Trp</td><td>Asn</td><td>Cheese</td><td>Gly</td><td>Ala</td><td>Leu</td><td>Thr</td><td>Cheese</td><td>Gly</td>

<td> 145</td><td></td><td></td><td></td><td></td><td> 150</td><td></td><td></td><td></td><td></td><td> 155</td><td></td><td></td><td></td><td></td><td> 160</td>

<td>Val</td><td>His</td><td>Thr</td><td>Phe</td><td>Pro</td><td>Ala</td><td>Val</td><td>Leu</td><td>Gin</td><td></td><td></td><td></td><td></td><td></td><td></td><td></td>

165 &lt;210 74 &lt;211&gt; 167 &lt;212&gt; PRT &lt;213&gt; Homo sapiens &lt;400 74

PL 214 003 Β1

<td>Gly 1</td><td>Val</td><td>Val</td><td>Gin</td><td>Pro 5</td><td>Gly</td><td>Arg</td><td>Cheese</td><td>Leu</td><td>Arg 10</td><td>Leu</td><td>Cheese</td><td>Cys</td><td>Val</td><td>Ala 15</td><td>Cheese</td>

<td>Gly</td><td>Phe</td><td>Thr</td><td>Phe</td><td>Cheese</td><td>Cheese</td><td>His</td><td>Gly</td><td>Underworld</td><td>His</td><td>Trp</td><td>Val</td><td>Arg</td><td>Gin</td><td>Ala</td><td>Pro</td>

<td></td><td></td><td></td><td> 20</td><td></td><td></td><td></td><td></td><td> 25</td><td></td><td></td><td></td><td></td><td> 30</td><td></td><td></td>

<td>Gly</td><td>Lys</td><td>Gly</td><td>Leu</td><td>Glu</td><td>Trp</td><td>Val</td><td>Ala</td><td>Val</td><td>How much</td><td>Trp</td><td>Tyr</td><td>Asp</td><td>Gly</td><td>Arg</td><td>Asn</td>

<td></td><td></td><td> 35</td><td></td><td></td><td></td><td></td><td> 40</td><td></td><td></td><td></td><td></td><td> 45</td><td></td><td></td><td></td>

<td>Lys</td><td>Tyr</td><td>Tyr</td><td>Ala</td><td>Asp</td><td>Cheese</td><td>Val</td><td>Lys</td><td>Gly</td><td>Arg</td><td>Phe</td><td>Thr</td><td>How much</td><td>Cheese</td><td>Arg</td><td>Asp</td>

<td></td><td> 50</td><td></td><td></td><td></td><td></td><td> 55</td><td></td><td></td><td></td><td></td><td> 60</td><td></td><td></td><td></td><td></td>

<td>Asn</td><td>Cheese</td><td>Lys</td><td>Asn</td><td>Thr</td><td>Leu</td><td>Phe</td><td>Leu</td><td>Gin</td><td>Underworld</td><td>Asn</td><td>Cheese</td><td>Leu</td><td>Arg</td><td>Ala</td><td>Glu</td>

<td> 65</td><td></td><td></td><td></td><td></td><td> 70</td><td></td><td></td><td></td><td></td><td> 75</td><td></td><td></td><td></td><td></td><td> 80</td>

<td>ASp</td><td>Thr</td><td>Ala</td><td>Val</td><td>Tyr</td><td>Tyr</td><td>Cys</td><td>Ala</td><td>Arg</td><td>Gly</td><td>Gly</td><td>His</td><td>Phe</td><td>Gly</td><td>Pro</td><td>Phe</td>

<td></td><td></td><td></td><td></td><td> 85</td><td></td><td></td><td></td><td></td><td> 90</td><td></td><td></td><td></td><td></td><td> 95</td><td></td>

<td>Asp</td><td>Tyr</td><td>Trp</td><td>Gly</td><td>Gin</td><td>Gly</td><td>Thr</td><td>Wow</td><td>Val</td><td>Thr</td><td>Val</td><td>Cheese</td><td>Cheese</td><td>Ala</td><td>Cheese</td><td>Thr</td>

<td></td><td></td><td></td><td> 100</td><td></td><td></td><td></td><td></td><td> 105</td><td></td><td></td><td></td><td></td><td> 110</td><td></td><td></td>

<td>Lys</td><td>Gly</td><td>Pro</td><td>Cheese</td><td>Val</td><td>Phe</td><td>Pro</td><td>Leu</td><td>Ala</td><td>Pro</td><td>cys</td><td>Cheese</td><td>Arg</td><td>Cheese</td><td>Thr</td><td>Cheese</td>

<td></td><td></td><td> 115</td><td></td><td></td><td></td><td></td><td> 120</td><td></td><td></td><td></td><td></td><td> 125</td><td></td><td></td><td></td>

<td>Glu</td><td>Cheese</td><td>Thr</td><td>Ala</td><td>Ala</td><td>Leu</td><td>Gly</td><td>Cys</td><td>Leu</td><td>Val</td><td>Lys</td><td>Asp</td><td>Tyr</td><td>Phe</td><td>Pro</td><td>Glu</td>

<td></td><td> 130</td><td></td><td></td><td></td><td></td><td> 135</td><td></td><td></td><td></td><td></td><td> 140</td><td></td><td></td><td></td><td></td>

<td>Pro</td><td>Val</td><td>Thr</td><td>val</td><td>Cheese</td><td>Trp</td><td>Asn</td><td>Cheese</td><td>Gly</td><td>Ala</td><td>Leu</td><td>Thr</td><td>Cheese</td><td>Gly</td><td>val</td><td>His</td>

<td> 145</td><td></td><td></td><td></td><td></td><td> 150</td><td></td><td></td><td></td><td></td><td> 155</td><td></td><td></td><td></td><td></td><td> 160</td>

<td>Thr</td><td>Phe</td><td>Pro</td><td>Ala</td><td>Val</td><td>Leu</td><td>Gin</td><td></td><td></td><td></td><td></td><td></td><td></td><td></td><td></td><td></td>

165 &lt;210&gt; 75 &lt;211&gt; 166 &lt;212&gt; PRT &lt;213&gt; Homo sapiens &lt;400&gt; 75

<td>Gly 1</td><td>Val</td><td>val</td><td>G .. n</td><td colspan="3">Pro Gly Arg 5</td><td>Cheese</td><td colspan="2">Leu Arg 10</td><td>Leu</td><td>Cheese</td><td>Cys</td><td>Thr</td><td>Ala 15</td><td>Cheese</td>

<td>Gly</td><td>Phe</td><td>Thr</td><td>Phe</td><td>Cheese</td><td>Asn</td><td>Tyr</td><td>dy</td><td>Underworld</td><td>His</td><td>Trp</td><td>Val</td><td>Arg</td><td>Gin</td><td>Ala</td><td>Pro</td>

<td></td><td></td><td></td><td> 20</td><td></td><td></td><td></td><td></td><td> 25</td><td></td><td></td><td></td><td></td><td> 30</td><td></td><td></td>

<td>Gly</td><td>Lys</td><td>Gly</td><td>Le u</td><td>Glu</td><td>Trp</td><td>Val</td><td>Ala</td><td>Val</td><td>How much</td><td>Trp</td><td>Tyr</td><td>Asp</td><td>Gly</td><td>Cheese</td><td>Asn</td>

<td></td><td></td><td> 35</td><td></td><td></td><td></td><td></td><td> 40</td><td></td><td></td><td></td><td></td><td> 45</td><td></td><td></td><td></td>

<td>Lys</td><td>His</td><td>Tyr</td><td>Gly</td><td>Asp</td><td>Cheese</td><td>Val</td><td>Lys</td><td>Gly</td><td>Arg</td><td>Phe</td><td>Thr</td><td>How much</td><td>Cheese</td><td>Cheese</td><td>Asp</td>

<td></td><td> 50</td><td></td><td></td><td></td><td></td><td> 55</td><td></td><td></td><td></td><td></td><td> 60</td><td></td><td></td><td></td><td></td>

<td>Asn</td><td>Cheese</td><td>Lys</td><td>Asn</td><td>Thr</td><td>Leu</td><td>Tyr</td><td>Leu</td><td>Gin</td><td>Underworld</td><td>Asn</td><td>Cheese</td><td>Leu</td><td>Arg</td><td>Ala</td><td>Glu</td>

<td> 65</td><td></td><td></td><td></td><td></td><td> 70</td><td></td><td></td><td></td><td></td><td> 75</td><td></td><td></td><td></td><td></td><td> 80</td>

Asp Thr Ala Val Tyr Tyr Cys Ala Arg Gly Glu Arg Leu Gly Ser Tyr

90 95

PL 214 003 Β1

<td>Phe</td><td>Asp</td><td>Tyr</td><td colspan="2">Trp Gly 100</td><td>Gin</td><td>Gly</td><td>Thr</td><td>Leu 105</td><td>Val</td><td>Thr</td><td>Val</td><td>Cheese</td><td>Cheese 110</td><td>Ala</td><td>Cheese</td>

<td>Thr</td><td>Lys</td><td>Gly</td><td>Pro</td><td>Cheese</td><td>Val</td><td>Phe</td><td>Pro</td><td>Leu</td><td>Ala</td><td>Pro</td><td>Cys</td><td>Cheese</td><td>Arg</td><td>Cheese</td><td>Thr</td>

<td></td><td></td><td> 115</td><td></td><td></td><td></td><td></td><td> 120</td><td></td><td></td><td></td><td></td><td> 12 5</td><td></td><td></td><td></td>

<td>Cheese</td><td>Glu</td><td>Cheese</td><td>Thr</td><td>Ala</td><td>Ala</td><td>Leu</td><td>Gly</td><td>Cys</td><td>Leu</td><td>Val</td><td>Lys</td><td>Asp</td><td>Tyr</td><td>Phe</td><td>Pro</td>

<td></td><td> 130</td><td></td><td></td><td></td><td></td><td> 135</td><td></td><td></td><td></td><td></td><td> 140</td><td></td><td></td><td></td><td></td>

<td>Glu</td><td>Pro</td><td>val</td><td>Thr</td><td>Val</td><td>Cheese</td><td>Trp</td><td>Asn</td><td>Cheese</td><td>Gly</td><td>Ala</td><td>Leu</td><td>Thr</td><td>Cheese</td><td>Gly</td><td>Val</td>

<td> 145</td><td></td><td></td><td></td><td></td><td> 150</td><td></td><td></td><td></td><td></td><td> 155</td><td></td><td></td><td></td><td></td><td> 160</td>

<td>His</td><td>Thr</td><td>Phe</td><td>Pro</td><td>Ala</td><td>Val</td><td></td><td></td><td></td><td></td><td></td><td></td><td></td><td></td><td></td><td></td>

165 &lt;210&gt; 76 &lt;211&gt; 167 &lt;212&gt; PRT &lt;213&gt; Homo sapiens

<td colspan="6"> &lt;400&gt; 76</td><td rowspan="2">Arg</td><td rowspan="2">Cheese</td><td colspan="2" rowspan="2">Leu Arg 10</td><td rowspan="2">Leu</td><td rowspan="2">Cheese</td><td rowspan="2">Cys</td><td rowspan="2">Val</td><td rowspan="2">Ala 15</td><td rowspan="2">Cheese</td>

<td>Gly 1</td><td>Val</td><td>Val</td><td>Gin</td><td>Pro 5</td><td>Gly</td>

<td>Gly</td><td>Phe</td><td>How much</td><td>Phe</td><td>Cheese</td><td>Cheese</td><td>His</td><td>Gly</td><td>How much</td><td>His</td><td>Trp</td><td>Val</td><td>Arg</td><td>Gin</td><td>Ala</td><td>Pro</td>

<td></td><td></td><td></td><td> 20</td><td></td><td></td><td></td><td></td><td> 25</td><td></td><td></td><td></td><td></td><td> 30</td><td></td><td></td>

<td>Gly</td><td>Lys</td><td>Gly</td><td>Leu</td><td>C-lu</td><td>Trp</td><td>Val</td><td>Ala</td><td>Val</td><td>How much</td><td>Trp</td><td>Tyr</td><td>Asp</td><td>Gly</td><td>Arg</td><td>Asn</td>

<td></td><td></td><td> 35</td><td></td><td></td><td></td><td></td><td> 40</td><td></td><td></td><td></td><td></td><td> 45</td><td></td><td></td><td></td>

<td>Lys</td><td>Asp</td><td>Tyr</td><td>Ala</td><td>ASp</td><td>Cheese</td><td>Val</td><td>Lys</td><td>Gly</td><td>Arg</td><td>Phe</td><td>Thr</td><td>How much</td><td>Cheese</td><td>Arg</td><td>Asp</td>

<td></td><td> 50</td><td></td><td></td><td></td><td></td><td> 55</td><td></td><td></td><td></td><td></td><td> 60</td><td></td><td></td><td></td><td></td>

<td>Asn</td><td>Cheese</td><td>lys</td><td>Asn</td><td>Thr</td><td>Leu</td><td>Tyr</td><td>Leu</td><td>Gin</td><td>Underworld</td><td>Asn</td><td>Cheese</td><td>Leu</td><td>Arg</td><td>Ala</td><td>Glu</td>

<td> 65</td><td></td><td></td><td></td><td></td><td> 70</td><td></td><td></td><td></td><td></td><td> 75</td><td></td><td></td><td></td><td></td><td> 80</td>

<td>Asp</td><td>Thr</td><td>Ala</td><td>val</td><td>Tyr</td><td>Tyr</td><td>Cys</td><td>Ala</td><td></td><td>Val</td><td>Ala</td><td>Pro</td><td>Leu</td><td>dy</td><td>Pro</td><td>Leu</td>

<td></td><td></td><td></td><td></td><td> 85</td><td></td><td></td><td></td><td></td><td> 90</td><td></td><td></td><td></td><td></td><td> 95</td><td></td>

<td>Asp</td><td>Tyr</td><td>Trp</td><td>G1 y</td><td>Gin</td><td>Giy</td><td>Thr</td><td>Leu</td><td>Val</td><td>Thr</td><td>Val</td><td>Cheese</td><td>Cheese</td><td>Al a</td><td>Cheese</td><td>Thr</td>

<td></td><td></td><td></td><td> 100</td><td></td><td></td><td></td><td></td><td> 105</td><td></td><td></td><td></td><td></td><td> 110</td><td></td><td></td>

<td>Lys</td><td>Gly</td><td>Pro</td><td>Cheese</td><td>Val</td><td>Phe</td><td>Pro</td><td>Leu</td><td>Ala</td><td>Pro</td><td>Cys</td><td>Cheese</td><td>Arg</td><td>Cheese</td><td>Thr</td><td>Cheese</td>

<td></td><td></td><td> 115</td><td></td><td></td><td></td><td></td><td> 120</td><td></td><td></td><td></td><td></td><td> 125</td><td></td><td></td><td></td>

<td>Glu</td><td>Cheese</td><td>Thr</td><td>Ala</td><td>Ala</td><td>Leu</td><td>Gly</td><td>Cys</td><td>Leu</td><td>Val</td><td>Lys</td><td>Asp</td><td>Tyr</td><td>Phe</td><td>Pro</td><td>Glu</td>

<td></td><td> 130</td><td></td><td></td><td></td><td></td><td> 135</td><td></td><td></td><td></td><td></td><td> 140</td><td></td><td></td><td></td><td></td>

<td>Pro</td><td>Val</td><td>Thr</td><td>Val</td><td>Cheese</td><td>Trp</td><td>Asn</td><td>Cheese</td><td>Gly</td><td>Ala</td><td>Leu</td><td>Thr</td><td>Cheese</td><td>Gly</td><td>Val</td><td>His</td>

<td> 145</td><td></td><td></td><td></td><td></td><td> 150</td><td></td><td></td><td></td><td></td><td> 155</td><td></td><td></td><td></td><td></td><td> 160</td>

Thr Phe Pro Ala Val Leu Gin &lt;210&gt; 77 &lt;211&gt; 153

PL 214 003 Β1 &lt;212&gt; PRT &lt;213&gt; Homo sapiens &lt;400&gt; 77

<td>Pro 1</td><td>Gly</td><td>Arg</td><td>Cheese</td><td>Leu 5</td><td>Arg</td><td>Leu</td><td>Cheese</td><td>Cys</td><td>Ala 10</td><td>But</td><td>Cheese</td><td>Gly</td><td>Phe</td><td>Thr 15</td><td>Phe</td>

<td>Cheese</td><td>Cheese</td><td>His</td><td>G1 y</td><td>How much</td><td>His</td><td>Trp</td><td>Val</td><td>Arg</td><td>Gin</td><td>Ala</td><td>Pro</td><td>Gly</td><td>Lys</td><td>Gly</td><td>Leu</td>

<td></td><td></td><td></td><td> 20</td><td></td><td></td><td></td><td></td><td> 25</td><td></td><td></td><td></td><td></td><td> 30</td><td></td><td></td>

<td>Glu</td><td>Trp</td><td>Val</td><td>Ala</td><td>Val</td><td>How much</td><td>Trp</td><td>Tyr</td><td>Asp</td><td>Gly</td><td>Arg</td><td>Asn</td><td>Lys</td><td>Asp</td><td>Tyr</td><td>Ala</td>

<td></td><td></td><td> 35</td><td></td><td></td><td></td><td></td><td> 40</td><td></td><td></td><td></td><td></td><td> 45</td><td></td><td></td><td></td>

<td>Asp</td><td>Cheese</td><td>Val</td><td>Lys</td><td>Gly</td><td>Arg</td><td>Phe</td><td>Thr</td><td>how much</td><td>Cheese</td><td>Arg</td><td>Asp</td><td>Asn</td><td>Cheese</td><td>Lys</td><td>Asn</td>

<td></td><td> 50</td><td></td><td></td><td></td><td></td><td> 55</td><td></td><td></td><td></td><td></td><td> 60</td><td></td><td></td><td></td><td></td>

<td>Thr</td><td>Leu</td><td>Tyr</td><td>Leu</td><td>Gin</td><td>Underworld</td><td>Asn</td><td>Cheese</td><td>Leu</td><td>Arg</td><td>Ala</td><td>Glu</td><td>Asp</td><td>Thr</td><td>Ala</td><td>Val</td>

<td> 65</td><td></td><td></td><td></td><td></td><td> 70</td><td></td><td></td><td></td><td></td><td> 75</td><td></td><td></td><td></td><td></td><td> 80</td>

<td>Tyr</td><td>Tyr</td><td>Cys</td><td>Ala</td><td>Arg</td><td>Val</td><td>Ala</td><td>Pro</td><td>Leu</td><td>Gly</td><td>Pro</td><td>Leu</td><td>Asp</td><td>Tyr</td><td>Trp</td><td>Gly</td>

<td></td><td></td><td></td><td></td><td> 85</td><td></td><td></td><td></td><td></td><td> 90</td><td></td><td></td><td></td><td></td><td> 95</td><td></td>

<td>Gin</td><td>Gly</td><td>Thr</td><td>Leu</td><td>Val</td><td>Thr</td><td>Val</td><td>Cheese</td><td>Cheese</td><td>Ala</td><td>Cheese</td><td>Thr</td><td>Lys</td><td>Gly</td><td>Pro</td><td>Cheese</td>

<td></td><td></td><td></td><td> 100</td><td></td><td></td><td></td><td></td><td> 105</td><td></td><td></td><td></td><td></td><td> 110</td><td></td><td></td>

<td>Val</td><td>Phe</td><td>Pro</td><td>Leu</td><td>Ala</td><td>Pro</td><td>Cys</td><td>Cheese</td><td>Arg</td><td>Cheese</td><td>Thr</td><td>Cheese</td><td>Glu</td><td>Cheese</td><td>Thr</td><td>Ala</td>

<td></td><td></td><td> 115</td><td></td><td></td><td></td><td></td><td> 120</td><td></td><td></td><td></td><td></td><td> 125</td><td></td><td></td><td></td>

<td>Ala</td><td>Leu</td><td>Gly</td><td>Cys</td><td>Leu</td><td>Val</td><td>Lys</td><td>Asp</td><td>Tyr</td><td>Phe</td><td>Pro</td><td>Glu</td><td>Pro</td><td>Val</td><td>Thr</td><td>Val</td>

<td></td><td> 130</td><td></td><td></td><td></td><td></td><td> 135</td><td></td><td></td><td></td><td></td><td> 140</td><td></td><td></td><td></td><td></td>

<td>Cheese</td><td>Trp</td><td>Asn</td><td>Cheese</td><td>Gly</td><td>Ala</td><td>Leu</td><td>Thr</td><td>Cheese</td><td></td><td></td><td></td><td></td><td></td><td></td><td></td>

145 150 &lt;210&gt; 78 &lt;211&gt; 163 &lt;212&gt; PRT &lt;213&gt; Homo sapiens &lt;400&gt; 78

<td>Pro 1.</td><td>Gly</td><td>Arg</td><td>Cheese</td><td colspan="2">Leu Arg 5</td><td>Leu</td><td>Cheese</td><td>Cys</td><td>Ala 10</td><td>Ara</td><td>Cheese</td><td>Gly</td><td>Phe</td><td>Thr 15</td><td>Phe</td>

<td>Cheese</td><td>Cheese</td><td>His</td><td>Gly</td><td>How much</td><td>His</td><td>Trp</td><td>Val</td><td>Arg</td><td>Gin</td><td>Ala</td><td>Pro</td><td>Gly</td><td>Lys</td><td>Gly</td><td>Leu</td>

<td></td><td></td><td></td><td> 20</td><td></td><td></td><td></td><td></td><td> 25</td><td></td><td></td><td></td><td></td><td> 30</td><td></td><td></td>

<td>Glu</td><td>Trp</td><td>Val</td><td>Ala</td><td>Val</td><td>How much</td><td>Trp</td><td>Tyr</td><td>Asp</td><td>Gly</td><td>Arg</td><td>Asn</td><td>Lys</td><td>Asp</td><td>Tyr</td><td>Ala</td>

<td></td><td></td><td> 35</td><td></td><td></td><td></td><td></td><td> 40</td><td></td><td></td><td></td><td></td><td> 45</td><td></td><td></td><td></td>

<td>Asp</td><td>Cheese</td><td>Val</td><td>Lys</td><td>Gly</td><td>Arg</td><td>Phe</td><td>Thr</td><td>How much</td><td>Cheese</td><td>Arg</td><td>Asp</td><td>Asn</td><td>Cheese</td><td>Lys</td><td>Lys</td>

<td></td><td> 50</td><td></td><td></td><td></td><td></td><td> 55</td><td></td><td></td><td></td><td></td><td> 60</td><td></td><td></td><td></td><td></td>

<td>Thr</td><td>Leu</td><td>Tyr</td><td>Leu</td><td>Gin</td><td>Underworld</td><td>Asn</td><td>Cheese</td><td>Le u</td><td>Arg</td><td>Ala</td><td>Glu</td><td>Asp</td><td>Thr</td><td>1 a</td><td>Val</td>

<td> 65</td><td></td><td></td><td></td><td></td><td> 70</td><td></td><td></td><td></td><td></td><td> 75</td><td></td><td></td><td></td><td></td><td> 80</td>

<td>Tyr</td><td>Tyr</td><td>Cys</td><td>Ala</td><td>Arg</td><td>Val</td><td>Ala</td><td>Pro</td><td>Leu</td><td>Gly</td><td>Pro</td><td>Leu</td><td>Asp</td><td>Tyr</td><td>Trp</td><td>Gly</td>

<td></td><td></td><td></td><td></td><td> 85</td><td></td><td></td><td></td><td></td><td> 90</td><td></td><td></td><td></td><td></td><td> 95</td><td></td>

PL 214 003 Β1

<td>Gin</td><td>Gly</td><td>Thr</td><td>Leu 100</td><td>Val</td><td>Thr</td><td>Val</td><td>Cheese</td><td>Cheese 105</td><td>Ala</td><td>Cheese</td><td>Thr</td><td>Lys</td><td>Gly 110</td><td>Pro</td><td>Cheese</td>

<td>Val</td><td>Phe</td><td>Pro</td><td>Leu</td><td>Ara</td><td>Pro</td><td>Cys</td><td>Cheese</td><td>Arg</td><td>Cheese</td><td>Thr</td><td>Cheese</td><td>Glu</td><td>Cheese</td><td>Thr</td><td>Ala</td>

<td></td><td></td><td> 115</td><td></td><td></td><td></td><td></td><td> 120</td><td></td><td></td><td></td><td></td><td> 125</td><td></td><td></td><td></td>

<td>Ala</td><td>Leu</td><td>Gly</td><td>Cys</td><td>Leu</td><td>vai</td><td>Lys</td><td>Asp</td><td>Tyr</td><td>Phe</td><td>Pro</td><td>Glu</td><td>Pro</td><td>Val</td><td>Thr</td><td>Val</td>

<td></td><td> 130</td><td></td><td></td><td></td><td></td><td> 135</td><td></td><td></td><td></td><td></td><td> 140</td><td></td><td></td><td></td><td></td>

<td>Cheese</td><td>Trp</td><td>Asn</td><td>Cheese</td><td>Gly</td><td>Ala</td><td>Leu</td><td>Thr</td><td>Cheese</td><td>Gly</td><td>Val</td><td>His</td><td>Thr</td><td>Phe</td><td>Pro</td><td>Ala</td>

<td> 145</td><td></td><td></td><td></td><td></td><td> 150</td><td></td><td></td><td></td><td></td><td> 155</td><td></td><td></td><td></td><td></td><td> 160</td>

<td>Val</td><td>Leu</td><td>Gin</td><td></td><td></td><td></td><td></td><td></td><td></td><td></td><td></td><td></td><td></td><td></td><td></td><td></td>

&lt;210&gt; 79 &lt;211&gt; 138 &lt;212&gt; PRT

<td colspan="3">&lt;213&gt; Homo</td><td colspan="2">sapiens</td><td></td><td></td><td></td><td></td><td></td><td></td><td></td><td></td><td></td><td></td><td></td>

<td colspan="3"> &lt;400 79</td><td></td><td></td><td></td><td></td><td></td><td></td><td></td><td></td><td></td><td></td><td></td><td></td><td></td>

<td>Gly</td><td>Gly</td><td>Val</td><td>Val</td><td>Glu</td><td>Pro</td><td>Giy</td><td>Arg</td><td>Cheese</td><td>Leu</td><td>Arg</td><td>Leu</td><td>Cheese</td><td>Cys</td><td>Thr</td><td>Ala</td>

<td> 1</td><td></td><td></td><td></td><td> 5</td><td></td><td></td><td></td><td></td><td> 10</td><td></td><td></td><td></td><td></td><td> 15</td><td></td>

<td>Cheese</td><td>Gly</td><td>Phe</td><td>Thr</td><td>Phe</td><td>Cheese</td><td>Cheese</td><td>Tyr</td><td>Gly</td><td>Underworld</td><td>His</td><td>Trp</td><td>Val</td><td>Arg</td><td>Gin</td><td>Ala</td>

<td></td><td></td><td></td><td> 20</td><td></td><td></td><td></td><td></td><td> 25</td><td></td><td></td><td></td><td></td><td> 30</td><td></td><td></td>

<td>Pro</td><td>Gly</td><td>Lys</td><td>Gly</td><td>Leu</td><td>Glu</td><td>Trp</td><td>Val</td><td>Ala</td><td>Val</td><td>How much</td><td>Trp</td><td>Tyr</td><td>Asp</td><td>Gly</td><td>Cheese</td>

<td></td><td></td><td> 35</td><td></td><td></td><td></td><td></td><td> 40</td><td></td><td></td><td></td><td></td><td> 45</td><td></td><td></td><td></td>

<td>Asn</td><td>Lys</td><td>His</td><td>Tyr</td><td>Al a</td><td>ASp</td><td>Cheese</td><td>Ala</td><td>Lys</td><td>Gly</td><td>Arg</td><td>Phe</td><td>Thr</td><td>How much</td><td>Cheese</td><td>Arg</td>

<td></td><td> 50</td><td></td><td></td><td></td><td></td><td> 55</td><td></td><td></td><td></td><td></td><td> 60</td><td></td><td></td><td></td><td></td>

<td>Asp</td><td>Asn</td><td>Cheese</td><td>Lys</td><td>Asn</td><td>Thr</td><td>Leu</td><td>Tyr</td><td>Leu</td><td>Gin</td><td>Underworld</td><td>Asn</td><td>Cheese</td><td>Leu</td><td>Arg</td><td>Ala</td>

<td> 65</td><td></td><td></td><td></td><td></td><td> 70</td><td></td><td></td><td></td><td></td><td> 75</td><td></td><td></td><td></td><td></td><td> 8 0</td>

<td>Glu</td><td>Asp</td><td>Thr</td><td>Ala</td><td>Val</td><td>Tyr</td><td>Tyr</td><td>Cys</td><td>Al a</td><td>Arg</td><td>Ala</td><td>Gly</td><td>Leu</td><td>Leu</td><td>dy</td><td>Tyr</td>

<td></td><td></td><td></td><td></td><td> 85</td><td></td><td></td><td></td><td></td><td> 90</td><td></td><td></td><td></td><td></td><td> 95</td><td></td>

<td>Phe</td><td>Asp</td><td>Tyr</td><td>Trp</td><td>Gly</td><td>Gin</td><td>Gly</td><td>Thr</td><td>Leu</td><td>val</td><td>Thr</td><td>Val</td><td>Cheese</td><td>Cheese</td><td>Ala</td><td>Cheese</td>

<td></td><td></td><td></td><td> 100</td><td></td><td></td><td></td><td></td><td> 105</td><td></td><td></td><td></td><td></td><td> 110</td><td></td><td></td>

<td>Thr</td><td>Lys</td><td>Gly</td><td>Pro</td><td>Cheese</td><td>Val</td><td>Phe</td><td>Pro</td><td>Leu</td><td>Ala</td><td>Pro</td><td>Cys</td><td>Cheese</td><td>Arg</td><td>Cheese</td><td>Thr</td>

<td></td><td></td><td> 115</td><td></td><td></td><td></td><td></td><td> 120</td><td></td><td></td><td></td><td></td><td> 125</td><td></td><td></td><td></td>

<td>Cheese</td><td>Glu</td><td>Cheese</td><td>Thr</td><td>Ala</td><td>Ala</td><td>Leu</td><td>Gly</td><td>Cys</td><td>Leu</td><td></td><td></td><td></td><td></td><td></td><td></td>

130 135 &lt;210&gt; 80 &lt;211&gt; 167 &lt;212&gt; PRT &lt;213&gt; Homo sapiens &lt;400&gt; 80

Gly Val Val Gin Pro Gly Arg Ser Leu Arg Leu Ser Cys Ala Ala Ser

5 10 15

PL 214 003 Β1

<td>dy</td><td>Phe</td><td>Thr</td><td>Phe twenty</td><td>Cheese</td><td>Cheese</td><td>Tyr</td><td colspan="2">Gly Met 25</td><td>His</td><td>Trp</td><td>Val</td><td>Arg</td><td>Gin thirty</td><td>Ala</td><td>Pro</td>

<td>Gly</td><td>Lys</td><td>dy</td><td>Leu</td><td>Glu</td><td>Trp</td><td>val</td><td>Ala</td><td>Val</td><td>How much</td><td>Trp</td><td>Tyr</td><td>Asp</td><td>Gly</td><td>Cheese</td><td>Asn</td>

<td></td><td></td><td> 35</td><td></td><td></td><td></td><td></td><td> 40</td><td></td><td></td><td></td><td></td><td> 45</td><td></td><td></td><td></td>

<td>Lys</td><td>Tyr</td><td>Tyr</td><td>Ara</td><td>Asp</td><td>Cheese</td><td>Val</td><td>Lys</td><td>Gly</td><td>Arg</td><td>Phe</td><td>Thr</td><td>How much</td><td>Cheese</td><td>Arg</td><td>Asp</td>

<td></td><td> 50</td><td></td><td></td><td></td><td></td><td> 55</td><td></td><td></td><td></td><td></td><td> 60</td><td></td><td></td><td></td><td></td>

<td>Asn</td><td>Cheese</td><td>Lys</td><td>Asn</td><td>Thr</td><td>Leu</td><td>Tyr</td><td>Leu</td><td>Gin</td><td>Underworld</td><td>Asn</td><td>Cheese</td><td>Leu</td><td>Arg</td><td>Ala</td><td>Glu</td>

<td> 65</td><td></td><td></td><td></td><td></td><td> 70</td><td></td><td></td><td></td><td></td><td> 75</td><td></td><td></td><td></td><td></td><td> 80</td>

<td>Asp</td><td>Thr</td><td>Ala</td><td>Val</td><td>Tyr</td><td>Tyr</td><td>Cys</td><td>Ala</td><td>Arg</td><td>Asp</td><td>Pro</td><td>Arg</td><td>Gly</td><td>Ala</td><td>Thr</td><td>Leu</td>

<td></td><td></td><td></td><td></td><td> 85</td><td></td><td></td><td></td><td></td><td> 90</td><td></td><td></td><td></td><td></td><td> 95</td><td></td>

<td>Tyr</td><td>Tyr</td><td>Tyr</td><td>Tyr</td><td>Tyr</td><td>dy</td><td>Underworld</td><td>Asp</td><td>Val</td><td>Trp</td><td>Gly</td><td>Gin</td><td>Gly</td><td>Thr</td><td>Thr</td><td>Val</td>

<td></td><td></td><td></td><td> 100</td><td></td><td></td><td></td><td></td><td> 105</td><td></td><td></td><td></td><td></td><td> 110</td><td></td><td></td>

<td>Thr</td><td>Val</td><td>Cheese</td><td>Cheese</td><td>Ala</td><td>Cheese</td><td>Thr</td><td>Lys</td><td>Gly</td><td>Pro</td><td>Cheese</td><td>Val</td><td>Phe</td><td>Pro</td><td>Leu</td><td>Al a</td>

<td></td><td></td><td> 115</td><td></td><td></td><td></td><td></td><td> 120</td><td></td><td></td><td></td><td></td><td> 125</td><td></td><td></td><td></td>

<td>Pro</td><td>Cys</td><td>Cheese</td><td>Arg</td><td>Cheese</td><td>Thr</td><td>Cheese</td><td>Glu</td><td>Cheese</td><td>Thr</td><td>Ala</td><td>Ala</td><td>Leu</td><td>Gly</td><td>Cys</td><td>Leu</td>

<td></td><td> 130</td><td></td><td></td><td></td><td></td><td> 135</td><td></td><td></td><td></td><td></td><td> 140</td><td></td><td></td><td></td><td></td>

<td>Val</td><td>Lys</td><td>Asp</td><td>Tyr</td><td>Phe</td><td>Pro</td><td>Glu</td><td>Pro</td><td>Val</td><td>Thr</td><td>Val</td><td>Cheese</td><td>Trp</td><td>Asn</td><td>Cheese</td><td>Gly</td>

<td> 1.45</td><td></td><td></td><td></td><td></td><td> 150</td><td></td><td></td><td></td><td></td><td> 155</td><td></td><td></td><td></td><td></td><td> 160</td>

<td>Ala</td><td>Leu</td><td>Thr</td><td>Cheese</td><td>Gly</td><td>Val</td><td>His</td><td></td><td></td><td></td><td></td><td></td><td></td><td></td><td></td><td></td>

165 &lt;210&gt; 81 &lt;2111 150 &lt;212&gt; PRT &lt;213&gt; Homo sapiens &lt;400 81

<td>Gly 1</td><td>V al</td><td>Val</td><td>Gin</td><td>Pro 5</td><td>Gly</td><td>Arg</td><td>Cheese</td><td>Leu</td><td>Arg r</td><td>Leu</td><td>Cheese</td><td>Cys</td><td>Ala</td><td>Ala 15</td><td>Cheese</td>

<td>Gly</td><td>Phe</td><td>Thr</td><td>Phe</td><td>Cheese</td><td>Cheese</td><td>Tyr</td><td>Gly</td><td>Underworld</td><td>His</td><td>Trp</td><td>Val</td><td>Arg</td><td>Gin</td><td>Ala</td><td>Pro</td>

<td></td><td></td><td></td><td> 20</td><td></td><td></td><td></td><td></td><td> 25</td><td></td><td></td><td></td><td></td><td> 30</td><td></td><td></td>

<td>dy</td><td>Lys</td><td>Gly</td><td>Leu</td><td>Glu</td><td>Trp</td><td>Val</td><td>Ala</td><td>Val</td><td>how much</td><td>Trp</td><td>Tyr</td><td>Asp</td><td>Gly</td><td>Cheese</td><td>His</td>

<td></td><td></td><td> 35</td><td></td><td></td><td></td><td></td><td> 40</td><td></td><td></td><td></td><td></td><td> 45</td><td></td><td></td><td></td>

<td>Lys</td><td>Tyr</td><td>Tyr</td><td>Ala</td><td>Asp</td><td>Cheese</td><td>Val</td><td>Lys</td><td>Gly</td><td>Arg</td><td>Phe</td><td>Thr</td><td>How much</td><td>Cheese</td><td>Arg</td><td>Asp</td>

<td></td><td> 50</td><td></td><td></td><td></td><td></td><td> 55</td><td></td><td></td><td></td><td></td><td> 60</td><td></td><td></td><td></td><td></td>

<td>Asn</td><td>Cheese</td><td>Lys</td><td>Asn</td><td>Thr</td><td>Leu</td><td>Tyr</td><td>Leu</td><td>Gin</td><td>Underworld</td><td>Asn</td><td>Cheese</td><td>Leu</td><td>Arg</td><td>Ala</td><td>Glu</td>

<td> 65</td><td></td><td></td><td></td><td></td><td> 70</td><td></td><td></td><td></td><td></td><td> 75</td><td></td><td></td><td></td><td></td><td> 80</td>

<td>Asp</td><td>Thr</td><td>Ala</td><td>Val</td><td>Tyr</td><td>Tyr</td><td>Cys</td><td>Ala</td><td>Arg</td><td>Gly</td><td>Ala</td><td>Val</td><td>Val</td><td>Val</td><td>Pro</td><td>Ala</td>

<td></td><td></td><td></td><td></td><td> 85</td><td></td><td></td><td></td><td></td><td> 90</td><td></td><td></td><td></td><td></td><td> 95</td><td></td>

<td>Ala</td><td>Underworld</td><td>Asp</td><td>Val</td><td>Ί'γ'ρ</td><td>Gly</td><td>Gin</td><td>Gly</td><td>Thr</td><td>Thr</td><td>Val</td><td>Thr</td><td>val</td><td>Cheese</td><td>Cheese</td><td>Ala</td>

<td></td><td></td><td></td><td> 100</td><td></td><td></td><td></td><td></td><td> 105</td><td></td><td></td><td></td><td></td><td> 110</td><td></td><td></td>

PL 214 003 Β1

<td>Cheese</td><td>Thr</td><td>Lys 115</td><td>Gly</td><td>Pro</td><td>Cheese</td><td>Val</td><td>Phe 120</td><td>Pro</td><td>Leu</td><td></td><td>Pro</td><td>Cys 125</td><td>Cheese</td><td>Arg</td><td>Cheese</td>

<td>Thr</td><td>Cheese</td><td>Glu</td><td>Cheese</td><td>Thr</td><td>Ala</td><td>Ala</td><td>Leu</td><td>Gly</td><td>cys</td><td>Leu</td><td>Val</td><td>cys</td><td>Asp</td><td>Tyr</td><td>Phe</td>

<td></td><td> 130</td><td></td><td></td><td></td><td></td><td> 135</td><td></td><td></td><td></td><td></td><td> 140</td><td></td><td></td><td></td><td></td>

<td>Pro</td><td>Glu</td><td>Pro</td><td>Val</td><td>Thr</td><td>Val</td><td></td><td></td><td></td><td></td><td></td><td></td><td></td><td></td><td></td><td></td>

145 150 &lt;210&gt; 82 &lt;211&gt; 146 &lt;212&gt; PRT

<td colspan="3">&lt;213&gt; Homo</td><td colspan="13">sapiens</td>

<td colspan="3"> &lt;400&gt; 82</td><td></td><td></td><td></td><td></td><td></td><td></td><td></td><td></td><td></td><td></td><td></td><td></td><td></td>

<td>Val</td><td>Val</td><td>Gin</td><td>Pro</td><td>Gly</td><td>Arg</td><td>Cheese</td><td>Leu</td><td>Arg</td><td>Leu</td><td>Cheese</td><td>Cys</td><td>Ala</td><td>Ala</td><td>Cheese</td><td>dy</td>

<td> 1</td><td></td><td></td><td></td><td>S.</td><td></td><td></td><td></td><td></td><td> 10</td><td></td><td></td><td></td><td></td><td> 15</td><td></td>

<td>Phe</td><td>Thr</td><td>Phe</td><td>Cheese</td><td>Cheese</td><td>Cys</td><td>dy</td><td>Underworld</td><td>His</td><td>Trp</td><td>Val</td><td>Arg</td><td>Gin</td><td>Ala</td><td>Pro</td><td>Gly</td>

<td></td><td></td><td></td><td> 20</td><td></td><td></td><td></td><td></td><td> 25</td><td></td><td></td><td></td><td></td><td> 30</td><td></td><td></td>

<td>Lys</td><td>Gly</td><td>Leu</td><td>Glu</td><td>Trp</td><td>Val</td><td>Ala</td><td>Val</td><td>How much</td><td>Trp</td><td>Cheese</td><td>Asp</td><td>Gly</td><td>Cheese</td><td>His</td><td>Lys</td>

<td></td><td></td><td> 35</td><td></td><td></td><td></td><td></td><td> 40</td><td></td><td></td><td></td><td></td><td> 45</td><td></td><td></td><td></td>

<td>Tyr</td><td>Tyr</td><td>Ala</td><td>Asp</td><td>Cheese</td><td>Val</td><td>Lys</td><td>Gly</td><td>Arg</td><td>Phe</td><td>Thr</td><td>How much</td><td>Cheese</td><td>Arg</td><td>Asp</td><td>Asn</td>

<td></td><td> 50</td><td></td><td></td><td></td><td></td><td> 55</td><td></td><td></td><td></td><td></td><td> 60</td><td></td><td></td><td></td><td></td>

<td>Cheese</td><td>Lys</td><td>Asn</td><td>Thr</td><td>Leu</td><td>Tyr</td><td>Leu</td><td>Gin</td><td>Underworld</td><td>Asn</td><td>Cheese</td><td>Leu</td><td>Arg</td><td>Ala</td><td>Glu</td><td>Asp</td>

<td> 65</td><td></td><td></td><td></td><td></td><td> 70</td><td></td><td></td><td></td><td></td><td> 75</td><td></td><td></td><td></td><td></td><td> 80</td>

<td>Thr</td><td>Al a</td><td>Val</td><td>Tyr</td><td>Tyr</td><td>Cys</td><td>Ala</td><td>Arg</td><td>Gly</td><td>Thr</td><td>Underworld</td><td>How much</td><td>Val</td><td>vai</td><td>Gly</td><td>Thr</td>

<td></td><td></td><td></td><td></td><td> 85</td><td></td><td></td><td></td><td></td><td> 90</td><td></td><td></td><td></td><td></td><td> 95</td><td></td>

<td>Leu</td><td>Asp</td><td>Tyr</td><td>Trp</td><td>Gly</td><td>Gin</td><td>Gly</td><td>Thr</td><td>Leu</td><td>Val</td><td>Thr</td><td>Val</td><td>Cheese</td><td>Cheese</td><td>Ala</td><td>Cheese</td>

<td></td><td></td><td></td><td> 100</td><td></td><td></td><td></td><td></td><td> 105</td><td></td><td></td><td></td><td></td><td> 110</td><td></td><td></td>

<td>Thr</td><td>Lys</td><td>Gly</td><td>Pro</td><td>Cheese</td><td>Val</td><td>Phe</td><td>Pro</td><td>Leu</td><td>Ala</td><td>Pro</td><td>cys</td><td>Cheese</td><td>Arg</td><td>Cheese</td><td>Thr</td>

<td></td><td></td><td> 115</td><td></td><td></td><td></td><td></td><td> 120</td><td></td><td></td><td></td><td></td><td> 125</td><td></td><td></td><td></td>

<td>Cheese</td><td>Gili</td><td>Cheese</td><td>Thr</td><td>Ala</td><td>Ala</td><td>Leu</td><td>Gly</td><td>Cys</td><td>Leu</td><td>Val</td><td>Lys</td><td>Asp</td><td>Tyr</td><td>Phe</td><td>Pro</td>

130 135 140

Glu Pro

<td> 145</td><td></td>

<td> &lt;210&gt;</td><td> 83</td>

<td> &lt;211&gt;</td><td> 171</td>

<td> &lt;212&gt;</td><td>PRT</td>

<td> &lt;213&gt;</td><td>Homo sapiens</td>

<td> &lt;400&gt;</td><td> 83</td>

<td>Gly 1</td><td>val</td><td>Val</td><td>Gin</td><td>Pro 5</td><td>Gly</td><td>Arg</td><td>Cheese</td><td>Leu</td><td>Arg 10</td><td>Leu</td><td>Cheese</td><td>Cys</td><td>Ala</td><td>Ala Ί C,</td><td>Cheese</td>

<td>Gly</td><td>Phe</td><td>Thr</td><td>Phe</td><td>Cheese</td><td>Cheese</td><td>Tyr</td><td>Gly</td><td>Val</td><td>His</td><td>Trp</td><td>Val</td><td>Arg</td><td>Gin</td><td>Ala</td><td>Pro</td>

PL 214 003 Β1

<td></td><td></td><td></td><td> 20</td><td></td><td></td><td></td><td></td><td> 25</td><td></td><td></td><td></td><td></td><td> 30</td><td></td><td></td>

<td>Gly</td><td>Lys</td><td>Gly</td><td>Leu</td><td>Glu</td><td>Trp</td><td>Val</td><td>Ala</td><td>Val</td><td>1 le</td><td>Trp</td><td>Tyr</td><td>Asp</td><td>Gly</td><td>Cheese</td><td>Asn</td>

<td></td><td></td><td> 35</td><td></td><td></td><td></td><td></td><td> 40</td><td></td><td></td><td></td><td></td><td> 45</td><td></td><td></td><td></td>

<td>Lys</td><td>Tyr</td><td>Tyr</td><td>Ala</td><td>Asp</td><td>Cheese</td><td>Val</td><td>Lys</td><td>C-ly</td><td>Arg</td><td>Phe</td><td>Thr</td><td>How much</td><td>Cheese</td><td>Arg</td><td>Asp</td>

<td></td><td> 50</td><td></td><td></td><td></td><td></td><td> 55</td><td></td><td></td><td></td><td></td><td> 60</td><td></td><td></td><td></td><td></td>

<td>Asn</td><td>Cheese</td><td>Lys</td><td>Cheese</td><td>Thr</td><td>Leu</td><td>Tyr</td><td>Leu</td><td>Gin</td><td>Underworld</td><td>Asn</td><td>Cheese</td><td>Leu</td><td>Arg</td><td>Ala</td><td>Glu</td>

<td> 65</td><td></td><td></td><td></td><td></td><td> 70</td><td></td><td></td><td></td><td></td><td> 75</td><td></td><td></td><td></td><td></td><td> 80</td>

<td>Asp</td><td>Thr</td><td>Ala</td><td>Val</td><td>Tyr</td><td>Tyr</td><td>Cys</td><td>Ala</td><td>Arg</td><td>Asp</td><td>Cheese</td><td>Tyr</td><td>Tyr</td><td>Asp</td><td>Phe</td><td>Trp</td>

<td></td><td></td><td></td><td></td><td> 85</td><td></td><td></td><td></td><td></td><td> 90</td><td></td><td></td><td></td><td></td><td> 95</td><td></td>

<td>Cheese</td><td>dy</td><td>Arg</td><td>Gly</td><td>Gly</td><td>Underworld</td><td>Asp</td><td>Val</td><td>Trp</td><td>Gly</td><td>Gin</td><td>Gly</td><td>Thr</td><td>Thr</td><td>Val</td><td>Thr</td>

<td></td><td></td><td></td><td> 100</td><td></td><td></td><td></td><td></td><td> 105</td><td></td><td></td><td></td><td></td><td> 110</td><td></td><td></td>

<td>Val</td><td>Cheese</td><td>Cheese</td><td>Ala</td><td>Cheese</td><td>Thr</td><td>Lys</td><td>Gly</td><td>Pro</td><td>Cheese</td><td>Val</td><td>Phe</td><td>Pro</td><td>Leu</td><td>Ala</td><td>Pro</td>

<td></td><td></td><td> 115</td><td></td><td></td><td></td><td></td><td> 120</td><td></td><td></td><td></td><td></td><td> 125</td><td></td><td></td><td></td>

<td>Cys</td><td>Cheese</td><td>Arg</td><td>Cheese</td><td>Thr</td><td>Cheese</td><td>Glu</td><td>Cheese</td><td>Thr</td><td>Ala</td><td>Ala</td><td>Leu</td><td>Gly</td><td>cys</td><td>Leu</td><td>Val</td>

<td></td><td> 130</td><td></td><td></td><td></td><td></td><td> 135</td><td></td><td></td><td></td><td></td><td> 140</td><td></td><td></td><td></td><td></td>

<td>Lys</td><td>Asp</td><td>Tyr</td><td>Phe</td><td>Pro</td><td>Glu</td><td>Pro</td><td>Val</td><td>Thr</td><td>Val</td><td>Cheese</td><td>Trp</td><td>Asn</td><td>Cheese</td><td>Gly</td><td>Ala</td>

<td> 145</td><td></td><td></td><td></td><td></td><td> 150</td><td></td><td></td><td></td><td></td><td> 155</td><td></td><td></td><td></td><td></td><td> 160</td>

<td>Leu</td><td>Thr</td><td>Cheese</td><td>Gly</td><td>val</td><td>His</td><td>Thr</td><td>Phe</td><td>Pro</td><td>Ala</td><td>Val</td><td></td><td></td><td></td><td></td><td></td>

165 170 &lt;210&gt; 84 &lt;211&gt; 163 &lt;212&gt; PRT &lt;213&gt; Homo sapiens &lt;4001 84

<td>Val 1</td><td>Gin</td><td>Pro</td><td>Gly</td><td>Arg 5</td><td>Cheese</td><td>Leu</td><td>Arg</td><td>Leu</td><td>Cheese 10</td><td>cys</td><td>Ala</td><td>Ala</td><td>Cheese</td><td>Gly 15</td><td>Phe</td>

<td>Thr</td><td>Phe</td><td>Cheese</td><td>Asn</td><td>Tyr</td><td>Ala</td><td>Underworld</td><td>His</td><td>Trp</td><td>Val</td><td>Arg</td><td>Gin</td><td>Ala</td><td>Pro</td><td>Gly</td><td>Lys</td>

<td></td><td></td><td></td><td> 20</td><td></td><td></td><td></td><td></td><td> 25</td><td></td><td></td><td></td><td></td><td> 30</td><td></td><td></td>

<td>Gly</td><td>Leu</td><td>Glu</td><td>Trp</td><td>Val</td><td>Val</td><td>Val</td><td>How much</td><td>Trp</td><td>His</td><td>Asp</td><td>dy</td><td>Asn</td><td>Asn</td><td>Lys</td><td>Tyr</td>

<td></td><td></td><td> 35</td><td></td><td></td><td></td><td></td><td> 40</td><td></td><td></td><td></td><td></td><td> 45</td><td></td><td></td><td></td>

<td>Tyr</td><td>Ala</td><td>Glu</td><td>Cheese</td><td>Val</td><td>Lys</td><td>Gly</td><td>Arg</td><td>Phe</td><td>Thr</td><td>How much</td><td>Cheese</td><td>Arg</td><td>Asp</td><td>Asn</td><td>Cheese</td>

<td></td><td> 50</td><td></td><td></td><td></td><td></td><td> 55</td><td></td><td></td><td></td><td></td><td> 60</td><td></td><td></td><td></td><td></td>

<td>Lys</td><td>Asn</td><td>Thr</td><td>Leu</td><td>Tyr</td><td>Leu</td><td>Gin</td><td>Underworld</td><td>Asn</td><td>Cheese</td><td>Leu</td><td>Arg</td><td>Ala</td><td>Glu</td><td>Asp</td><td>Thr</td>

<td> 65</td><td></td><td></td><td></td><td></td><td> 70</td><td></td><td></td><td></td><td></td><td> 75</td><td></td><td></td><td></td><td></td><td> 80</td>

<td>Ala</td><td>Val</td><td>Tyr</td><td>Tyr</td><td>Cys</td><td>Ala</td><td>Arg</td><td>Asp</td><td>Gin</td><td>Gly</td><td>Thr</td><td>Gly</td><td>Trp</td><td>Tyr</td><td>Gly</td><td>Gly</td>

<td></td><td></td><td></td><td></td><td> 85</td><td></td><td></td><td></td><td></td><td> 90</td><td></td><td></td><td></td><td></td><td> 95</td><td></td>

<td>Phe</td><td>Asp</td><td>Phe</td><td>Trp</td><td>Gly</td><td>Gin</td><td>Gly</td><td>Thr</td><td>Leu</td><td>Va.l</td><td>Thr</td><td>Val</td><td>Cheese</td><td>Cheese</td><td>Ala</td><td>Cheese</td>

<td></td><td></td><td></td><td> 100</td><td></td><td></td><td></td><td></td><td> 105</td><td></td><td></td><td></td><td></td><td> 110</td><td></td><td></td>

Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Cys Ser Arg Ser Thr

PL 214 003 Β1

<td></td><td></td><td> 115</td><td></td><td></td><td></td><td></td><td> 120</td><td></td><td></td><td></td><td></td><td> 125</td><td></td><td></td><td></td>

<td>Cheese</td><td>Glu</td><td>Cheese</td><td>Thr</td><td>Ala</td><td>Ala</td><td>Leu</td><td>Gly</td><td>Cys</td><td>Leu</td><td>Val</td><td>Lys</td><td>Asp</td><td>Tyr</td><td>Ehe</td><td>Pro</td>

<td></td><td> 130</td><td></td><td></td><td></td><td></td><td> 135</td><td></td><td></td><td></td><td></td><td> 140</td><td></td><td></td><td></td><td></td>

<td>-Glu</td><td>Pro</td><td>Val</td><td>Thr</td><td>Val</td><td>Cheese</td><td>Trp</td><td>Asn</td><td>Cheese</td><td>Gly</td><td>Ala</td><td>Leu</td><td>Thr</td><td>Cheese</td><td>Gly</td><td>Val</td>

<td> 145</td><td></td><td></td><td></td><td></td><td> 150</td><td></td><td></td><td></td><td></td><td> 155</td><td></td><td></td><td></td><td></td><td> 160</td>

His Thr Phe &lt;210&gt; 85 &lt;211&gt; 76 &lt;212&gt; PRT &lt;213&gt; Homo sapiens

<td colspan="6"> &lt;400&gt; 85</td><td rowspan="2">Cheese</td><td colspan="2" rowspan="2">Gly cheese</td><td rowspan="2">Gly 10</td><td rowspan="2">Tyr</td><td rowspan="2">Tyr</td><td rowspan="2">• rrp</td><td rowspan="2">Cheese</td><td rowspan="2">Trp 15</td><td rowspan="2">How much</td>

<td>Val 1</td><td>Cheese</td><td>Gly</td><td colspan="2">Gly Ser 5</td><td>How much</td>

<td>Arg</td><td>Gin</td><td>His</td><td>Pro</td><td>Gly</td><td>Lys</td><td>Gly</td><td>Leu</td><td>Glu</td><td>Trp</td><td>How much</td><td>Gly</td><td>Tyr</td><td>How much</td><td>Tyr</td><td>Tyr</td>

<td></td><td></td><td></td><td> 20</td><td></td><td></td><td></td><td></td><td> 25</td><td></td><td></td><td></td><td></td><td> 30</td><td></td><td></td>

<td>Cheese</td><td>Gly</td><td>Cheese</td><td>Thr</td><td>Tyr</td><td>Tyr</td><td>Asn</td><td>Pro</td><td>Cheese</td><td>Leu</td><td>Lys</td><td>Cheese</td><td>Arg</td><td>Val</td><td>Th from?</td><td>How much</td>

<td></td><td></td><td> 35</td><td></td><td></td><td></td><td></td><td> 40</td><td></td><td></td><td></td><td></td><td> 45</td><td></td><td></td><td></td>

<td>Cheese</td><td>Val</td><td>Asp</td><td>Thr</td><td>Cheese</td><td>Iiys</td><td>Asn</td><td>Gin</td><td>Phe</td><td>Cheese</td><td>Leu</td><td>Lys</td><td>Leu</td><td>Cheese</td><td>Cheese</td><td>Val</td>

<td></td><td> 50</td><td></td><td></td><td></td><td></td><td> 55</td><td></td><td></td><td></td><td></td><td> 60</td><td></td><td></td><td></td><td></td>

<td>Thr</td><td>Ala</td><td>Ala</td><td>Asp</td><td>Thr</td><td>Ala</td><td>Val</td><td>Tyr</td><td>Tyr</td><td>Cys</td><td>Ala</td><td>Arg</td><td></td><td></td><td></td><td></td>

70 75 &lt;210&gt; 86 &lt;211&gt; 172.

&lt;212&gt; PRT &lt;213&gt; Homo sapiens

<td colspan="3"> &lt;400&gt; 86</td><td rowspan="2">Gly</td><td rowspan="2">Leu 5</td><td rowspan="2">Val</td><td rowspan="2">Lys</td><td rowspan="2">Pro</td>

<td>Cheese 1</td><td>Gly</td><td>Pro</td>

<td>Thr</td><td>Val</td><td>Cheese</td><td>Gly twenty</td><td>Gly</td><td>Cheese</td><td>How much</td><td>Cheese</td>

<td>How much</td><td>Arg</td><td>Gin 35</td><td>His</td><td>Pro</td><td>Gly</td><td>Lys</td><td>Gly 40</td>

<td>Tyr</td><td>How much 50</td><td>Gly</td><td>Asn</td><td>Thr</td><td>Tyr</td><td>Tyr 55</td><td>Asn</td>

<td>How much 65</td><td>Cheese</td><td>Val</td><td>Asp</td><td>Thr</td><td>Cheese 70</td><td>Lys</td><td>Asn</td>

<td>Val</td><td>Thr</td><td>AT a</td><td>Ala</td><td>Asp 85</td><td>Thr</td><td>Ala</td><td>Val</td>

<td></td><td>Gin 10</td><td>How much</td><td>Leu</td><td>Cheese</td><td>Leu</td><td>Thr 15</td><td>cys</td>

<td>Cheese 25</td><td>Gly</td><td>Gly</td><td>His</td><td>Tyr</td><td>Trp thirty</td><td>Cheese</td><td>Trp</td>

<td>Leu</td><td>Glu</td><td>Trp</td><td>Background</td><td>Gly 45</td><td>Tyr</td><td>How much</td><td>Tyr</td>

<td>Pro</td><td>Cheese</td><td>Leu</td><td>Lys 60</td><td>Cheese</td><td>Arg</td><td>Val</td><td>Thr</td>

<td>Gin</td><td>Phe</td><td>Cheese 75</td><td>Leu</td><td>Lys</td><td>Leu</td><td>Cheese</td><td>Cheese 80</td>

<td>Tyr</td><td>Tyr 90</td><td>Cys</td><td>Ala</td><td>Arg</td><td>Asp</td><td>Cheese 95</td><td>Gly</td>

Asp Tyr Tyr Gly Ile Asp Val Trp Gly Gin Gly Thr Thr Val Thr Val

PL 214 003 Β1

<td></td><td></td><td></td><td> 100</td><td></td><td></td><td></td><td></td><td> 105</td><td></td><td></td><td></td><td></td><td> 110</td><td></td><td></td>

<td>Cheese</td><td>Cheese</td><td>Ala</td><td>Cheese</td><td>ΈΙίχ</td><td>Lys</td><td>Gly</td><td>Pro</td><td>Cheese</td><td>Val</td><td>Phe</td><td>Pro</td><td>Leu</td><td>Ala</td><td>Pro</td><td>Cys</td>

<td></td><td></td><td> 115</td><td></td><td></td><td></td><td></td><td> 120</td><td></td><td></td><td></td><td></td><td> 125</td><td></td><td></td><td></td>

<td>Cheese</td><td>Arg</td><td>Cheese</td><td>Thr</td><td>Cheese</td><td>Glu</td><td>Cheese</td><td>Thr</td><td>Ala</td><td>Ala</td><td>Leu</td><td>Gly</td><td>Cys</td><td>Leu</td><td>Val</td><td>Lys</td>

<td></td><td> 130</td><td></td><td></td><td></td><td></td><td> 135</td><td></td><td></td><td></td><td></td><td> 140</td><td></td><td></td><td></td><td></td>

<td>Asp</td><td>Tyr</td><td>Phe</td><td>Pro</td><td>Glu</td><td>Pro</td><td>Val</td><td>Thr</td><td>Val</td><td>Cheese</td><td>Trp</td><td>Asn</td><td>Cheese</td><td>Gly</td><td>Ala</td><td>Leu</td>

<td> 145</td><td></td><td></td><td></td><td></td><td> 150</td><td></td><td></td><td></td><td></td><td> 155</td><td></td><td></td><td></td><td></td><td> 160</td>

<td>Thr</td><td>Cheese</td><td>Gly</td><td>Val</td><td>His</td><td>Thr</td><td>Phe</td><td>Pro</td><td>Ala</td><td>Val</td><td>Leu</td><td>Gin</td><td></td><td></td><td></td><td></td>

165 170 &lt;210&gt; 87 &lt;211&gt; 96 &lt;212&gt; PRT &lt;213&gt; Homo sapiens

<td colspan="16"> &lt;400&gt; 87</td>

<td>Glu 1</td><td colspan="2">How many Val</td><td>Leu</td><td>Thr 5</td><td>Gin</td><td>Cheese</td><td>Pro</td><td>Gly</td><td>Thr 10</td><td>Leu</td><td>Cheese</td><td>Leu</td><td>Cheese</td><td>Pro 15</td><td>dy</td>

<td>Glu</td><td>Arg</td><td>Ala</td><td>Thr</td><td>Leu</td><td>Cheese</td><td>Cys</td><td>Arg</td><td>Ala</td><td>Cheese</td><td>Gin</td><td>Cheese</td><td>Val</td><td>Cheese</td><td>Cheese</td><td>Cheese</td>

<td></td><td></td><td></td><td> 2 0</td><td></td><td></td><td></td><td></td><td> 25</td><td></td><td></td><td></td><td></td><td> 30</td><td></td><td></td>

<td>Tyr</td><td>Leu</td><td>Ala</td><td>Trp</td><td>Tyr</td><td>Gin</td><td>Gin</td><td>Lys</td><td>Pro</td><td>Gly</td><td>Gin</td><td>Ala</td><td>Pro</td><td>Arg</td><td>Leu</td><td>Leu</td>

<td></td><td></td><td> 35</td><td></td><td></td><td></td><td></td><td> 40</td><td></td><td></td><td></td><td></td><td> 45</td><td></td><td></td><td></td>

<td>How much</td><td>Tyr</td><td>Gly</td><td>Ala</td><td>Cheese</td><td>Cheese</td><td>Arg</td><td>Ala</td><td>Thr</td><td>Gly</td><td>how much</td><td>Pro</td><td>Asp</td><td>Arg</td><td>Phe</td><td>Cheese</td>

<td></td><td> 50</td><td></td><td></td><td></td><td></td><td> 55</td><td></td><td></td><td></td><td></td><td> 60</td><td></td><td></td><td></td><td></td>

<td>Gly</td><td>Cheese</td><td>Gly</td><td>Cheese</td><td>Gly</td><td>Thr</td><td>Asp</td><td>Phe</td><td>Thr</td><td>Leu</td><td>Thr</td><td>How much</td><td>Cheese</td><td>Arg</td><td>Leu</td><td>Glu</td>

<td> 65</td><td></td><td></td><td></td><td></td><td> 70</td><td></td><td></td><td></td><td></td><td> 75</td><td></td><td></td><td></td><td></td><td>Θ0</td>

<td>Pro</td><td>Glu</td><td>Asp</td><td>Phe</td><td>Ais</td><td>Val</td><td>Tyr</td><td>Tyr</td><td>Cys</td><td>Gin</td><td>Gin</td><td>Tyr</td><td>Gly</td><td>Cheese</td><td>Cheese</td><td>Pro</td>

<td></td><td></td><td></td><td></td><td> 85</td><td></td><td></td><td></td><td></td><td> 90</td><td></td><td></td><td></td><td></td><td> 95</td><td></td>

&lt;210&gt; 88 &lt;211&gt; 141 &lt;212&gt; PRT &lt;213&gt; Homo sapiens

<td colspan="7"> &lt;400&gt; 88</td><td rowspan="2">Leu</td><td rowspan="2">Cheese</td><td rowspan="2">Pro 10</td><td rowspan="2">Gly</td><td rowspan="2">Glu</td><td rowspan="2">Arg</td><td rowspan="2">Ala</td><td rowspan="2">Thr 15</td><td rowspan="2">Leu</td>

<td>Gin 1</td><td>Cheese</td><td colspan="2">Pro Gly</td><td>Thr 5</td><td>Leu</td><td>Cheese</td>

<td>Cheese</td><td>Cys</td><td>Arg</td><td>Ala twenty</td><td>Cheese</td><td>Gin</td><td>Cheese</td><td>How much</td><td>Cheese 25</td><td>Cheese</td><td>Cheese</td><td>Phe</td><td>Leu</td><td>Al 3 thirty</td><td>Trp</td><td>Tyr</td>

<td>Gin</td><td>Gin</td><td>Arg 35</td><td>Pro</td><td>Gly</td><td>Gin</td><td>Ala</td><td>Pro 40</td><td>Arg</td><td>Leu</td><td>Leu</td><td> 11^</td><td>Tyr 45</td><td>Gly</td><td>Ala</td><td>Cheese</td>

<td>Cheese</td><td>Arg</td><td>Ala</td><td>Thr</td><td>Gly</td><td>How much</td><td>Pro</td><td>Asp</td><td>Arg</td><td>Phe</td><td>Cheese</td><td>Gly</td><td>Cheese</td><td>Gly</td><td>Cheese</td><td>Gly</td>

55 60

100

PL 214 003 Β1

<td>Thr 65</td><td>Asp</td><td>Phe</td><td>Thr</td><td>Leu</td><td>Thr 0</td><td>How much</td><td>Cheese</td><td>Arg</td><td>Leu</td><td>Glu 75</td><td>Pro</td><td>Glu</td><td>Asp</td><td>Phe</td><td>Al 3 SC</td>

<td>Val</td><td>Tyr</td><td>Tyr</td><td>Cys</td><td>Gin 35</td><td>Gin</td><td>Tyr</td><td>Gly</td><td>Thr</td><td>Cheese 90</td><td>Pro</td><td>Trp</td><td>Thr</td><td>Phe</td><td>Gly 95</td><td>Gin</td>

<td>Gly</td><td>Thr</td><td>Lys</td><td>Val 100</td><td>Glu</td><td>How much</td><td>Lys</td><td>Arg</td><td>Thr 105</td><td>Val</td><td>Ala</td><td>Ala</td><td>Pro</td><td>Cheese 110</td><td>Val</td><td>Phe</td>

<td>How much</td><td>Phe</td><td>Pro 115</td><td>Pro</td><td>Cheese</td><td>Asp</td><td>Glu</td><td>Gin 120</td><td>Leu</td><td>Lys</td><td>Cheese</td><td>Gly</td><td>Thr 125</td><td>Ala</td><td>Cheese</td><td>Val</td>

<td>Val</td><td>Cys</td><td>Leu</td><td>Leu</td><td>Asn</td><td>Asn</td><td>Phe</td><td>Tyr</td><td>Pro</td><td>Arg</td><td>Glu</td><td> 3</td><td>Lys</td><td></td><td></td><td></td>

130 135 140 &lt;210&gt; 89 &lt;211&gt; 141 &lt;212&gt; PRT &lt;213&gt; Homo sapiens &lt;400&gt; 89

<td>Gin 1</td><td>Cheese</td><td>Pro</td><td>Gly</td><td colspan="2">Thr Leu 5</td><td>Cheese</td><td>Leu</td><td>Cheese</td><td>Pro 10</td><td>Gly</td><td>Glu</td><td>Arg</td><td>Ala</td><td>Thr 15</td><td>Leu</td>

<td>Cheese</td><td>Cys</td><td>Arg</td><td>Thr</td><td>Cheese</td><td>Val</td><td>Cheese</td><td>Cheese</td><td>Cheese</td><td>Tyr</td><td>Leu</td><td>Ala</td><td>Trp</td><td>Tyr</td><td>Gin</td><td>Gin</td>

<td></td><td></td><td></td><td> 20</td><td></td><td></td><td></td><td></td><td> 25</td><td></td><td></td><td></td><td></td><td> 30</td><td></td><td></td>

<td>Lys</td><td>Pro</td><td>Gly</td><td>Gin</td><td>Ala</td><td>Pro</td><td>Arg</td><td>Leu</td><td>Leu</td><td>How much</td><td>Tyr</td><td>Gly</td><td>Ala</td><td>Cheese</td><td>Cheese</td><td>Arg</td>

<td></td><td></td><td> 35</td><td></td><td></td><td></td><td></td><td> 40</td><td></td><td></td><td></td><td></td><td> 45</td><td></td><td></td><td></td>

<td>Ala</td><td>Thr</td><td>Gly</td><td>How much</td><td>Pro</td><td>Asp</td><td>Arg</td><td>Phe</td><td>Cheese</td><td>Gly</td><td>Cheese</td><td>Gly</td><td>Cheese</td><td>Gly</td><td>Thr</td><td>Asp</td>

<td></td><td> 50</td><td></td><td></td><td></td><td></td><td> 55</td><td></td><td></td><td></td><td></td><td> 60</td><td></td><td></td><td></td><td></td>

<td>Phe</td><td>Thr</td><td>Leu</td><td>Thr</td><td>How much</td><td>Cheese</td><td>Arg</td><td>Leu</td><td>Glu</td><td>Pro</td><td>Glu</td><td>Asp</td><td>Phe</td><td>Al a</td><td>Val</td><td>Tyr</td>

<td> 65</td><td></td><td></td><td></td><td></td><td> 70</td><td></td><td></td><td></td><td></td><td> 75</td><td></td><td></td><td></td><td></td><td> 80</td>

<td>Tyr</td><td>Cys</td><td>Gin</td><td>Gin</td><td>Tyr</td><td>Gly</td><td>How much</td><td>Cheese</td><td>Pro</td><td>Phe</td><td>Thr</td><td>Phe</td><td>Gly</td><td>Gly</td><td>Gly</td><td>Thr</td>

<td></td><td></td><td></td><td></td><td> 85</td><td></td><td></td><td></td><td></td><td> 90</td><td></td><td></td><td></td><td></td><td> 95</td><td></td>

<td>Lys</td><td>Val</td><td>Glu</td><td>How much</td><td>Lys</td><td>Arg</td><td>Thr</td><td>Val</td><td>Ala</td><td>Ala</td><td>Pro</td><td>Cheese</td><td>Va, l</td><td>Phe</td><td>How much</td><td>Phe</td>

<td></td><td></td><td></td><td> 100</td><td></td><td></td><td></td><td></td><td> 105</td><td></td><td></td><td></td><td></td><td> 110</td><td></td><td></td>

<td>Pro</td><td>Pro</td><td>Cheese</td><td>Asp</td><td>Glu</td><td>Gin</td><td>Leu</td><td>Lys</td><td>Cheese</td><td>Gly</td><td>Thr</td><td>Ala</td><td>Cheese</td><td>Val</td><td>Val</td><td>Cys</td>

<td></td><td></td><td> 115</td><td></td><td></td><td></td><td></td><td> 120</td><td></td><td></td><td></td><td></td><td> 125</td><td></td><td></td><td></td>

<td>Leu</td><td>Leu</td><td>Asn</td><td>Asn</td><td>Phe</td><td>Tyr</td><td>Pro</td><td>Arg</td><td>Glu</td><td>Ala</td><td>Lys</td><td>Val</td><td>Gin</td><td></td><td></td><td></td>

<td></td><td> 130</td><td></td><td></td><td></td><td></td><td> 135</td><td></td><td></td><td></td><td></td><td> 140</td><td></td><td></td><td></td><td></td>

&lt;210&gt; 90 &lt;211&gt; 139 &lt;212&gt; PRT &lt;213&gt; Homo sapiens &lt;400&gt; 90

Gly Thr Leu Ser Leu Ser Pro Gly Glu Arg Ala Thr Leu Ser Cys Arg 1 5 10 15

PL 214 003 Β1

101

<td>Al 3</td><td>Cheese</td><td>Gin</td><td>Cheese twenty</td><td>Val</td><td>Cheese</td><td>Cheese</td><td>Tyr</td><td>Leu 25</td><td>Ala</td><td>Trp</td><td>iyr</td><td>Gin</td><td>Gin thirty</td><td>Lys</td><td>Pro</td>

<td>Gly</td><td>Gin</td><td>Ala</td><td>Pro</td><td>Arg</td><td>Leu</td><td>Leu</td><td>How much</td><td>Tyr</td><td>Gly</td><td>Ala</td><td>Cheese</td><td>Cheese</td><td>Arg</td><td>Al a</td><td>Thr</td>

<td></td><td></td><td> 35</td><td></td><td></td><td></td><td></td><td> 40</td><td></td><td></td><td></td><td></td><td> 45</td><td></td><td></td><td></td>

<td>G1 y</td><td>Background</td><td>Pro</td><td>Asp</td><td>Arg</td><td>Phe</td><td>Cheese</td><td>Gly</td><td>Cheese</td><td>Gly</td><td>Cheese</td><td>Gly</td><td>Thr</td><td>Asp</td><td>Phe</td><td>Thr</td>

<td></td><td> 50</td><td></td><td></td><td></td><td></td><td> 55</td><td></td><td></td><td></td><td></td><td> 60</td><td></td><td></td><td></td><td></td>

<td>Leu</td><td>Thr</td><td>How much</td><td>Cheese</td><td>Arg</td><td>Flax</td><td>Glu</td><td>Pro</td><td>Glu</td><td>Asp</td><td>Phe</td><td>Ala</td><td>val</td><td>Tyr</td><td>Tyr</td><td>Cys</td>

<td> 65</td><td></td><td></td><td></td><td></td><td> 70</td><td></td><td></td><td></td><td></td><td> 75</td><td></td><td></td><td></td><td></td><td> 80</td>

<td>Gin</td><td>Gin</td><td>Tyr</td><td>Gly</td><td>Arg</td><td>Cheese</td><td>Pro</td><td>Phe</td><td>Thr</td><td>Phe</td><td>Gly</td><td>Pro</td><td>Gly</td><td>Thr</td><td>Lys</td><td>Val</td>

<td></td><td></td><td></td><td></td><td> 8 5</td><td></td><td></td><td></td><td></td><td> 90</td><td></td><td></td><td></td><td></td><td> 95</td><td></td>

<td>Asp</td><td>How much</td><td>Lys</td><td>Arg</td><td>Thr</td><td>vai</td><td>Ala</td><td>Ala</td><td>Pro</td><td>Cheese</td><td>Val</td><td>Phe</td><td>I ie</td><td>Phe</td><td>Pro</td><td>Pro</td>

<td></td><td></td><td></td><td> 100</td><td></td><td></td><td></td><td></td><td> 105</td><td></td><td></td><td></td><td></td><td> 110</td><td></td><td></td>

<td>Cheese</td><td>Asp</td><td>Glu</td><td>Gin</td><td>Leu</td><td>Lys</td><td>Cheese</td><td>Gly</td><td>Thr</td><td>Ala</td><td>Cheese</td><td>Val</td><td>Val</td><td>Cys</td><td>Leu</td><td>Leu</td>

<td></td><td></td><td> 115</td><td></td><td></td><td></td><td></td><td> 120</td><td></td><td></td><td></td><td></td><td> 125</td><td></td><td></td><td></td>

<td>Asn</td><td>Asn</td><td>Phe</td><td>Tyr</td><td>Pro</td><td>Arg</td><td>Glu</td><td>Ala</td><td>Lys</td><td>Val</td><td>Gin</td><td></td><td></td><td></td><td></td><td></td>

130 135 &lt;210&gt; 91 &lt;211&gt; 142 &lt;212&gt; PRT &lt;213&gt; Homo sapiens &lt;400&gt; 91

<td>Gin 1</td><td>Cheese</td><td>Pro</td><td colspan="2">Gly Thr 5</td><td>Leu</td><td>Cheese</td><td>Leu</td><td>Cheese</td><td>Pro 10</td><td>Gly</td><td>Glu</td><td>Arg</td><td>Ala</td><td>Thr 15</td><td>Leu</td>

<td>Cheese</td><td>Cys</td><td>Arg</td><td>Ala</td><td>Cheese</td><td>Gin</td><td>Cheese</td><td>Val</td><td>Cheese</td><td>Cheese</td><td>Tyr</td><td>Leu</td><td>Ala</td><td>Trp</td><td>Tyr</td><td>Gin</td>

<td></td><td></td><td></td><td> 20</td><td></td><td></td><td></td><td></td><td> 25</td><td></td><td></td><td></td><td></td><td> 30</td><td></td><td></td>

<td>Gin</td><td>Lys</td><td>Pro</td><td>Giy</td><td>Gin</td><td>Ala</td><td>Pro</td><td>Arg</td><td>Pro</td><td>Leu</td><td>How much</td><td>Tyr</td><td>Gly</td><td>Val</td><td>Cheese</td><td>Cheese</td>

<td></td><td></td><td> 35</td><td></td><td></td><td></td><td></td><td> 40</td><td></td><td></td><td></td><td></td><td> 45</td><td></td><td></td><td></td>

<td>Arg</td><td>Ala</td><td>Thr</td><td>el y</td><td>How much</td><td>Pro</td><td>Asp</td><td>Arg</td><td>Phe</td><td>Cheese</td><td>Gly</td><td>Cheese</td><td>dy</td><td>Cheese</td><td>Gly</td><td>Thr</td>

<td></td><td> 50</td><td></td><td></td><td></td><td></td><td> 55</td><td></td><td></td><td></td><td></td><td> 60</td><td></td><td></td><td></td><td></td>

<td>Asp</td><td>Phe</td><td>Thr</td><td>Leu</td><td>Thr</td><td>How much</td><td>Cheese</td><td>Arg</td><td>Leu</td><td>Glu</td><td>Pro</td><td>Glu</td><td>Asp</td><td>Phe</td><td>Ala</td><td>Val</td>

<td> 65</td><td></td><td></td><td></td><td></td><td> 70</td><td></td><td></td><td></td><td></td><td> 75</td><td></td><td></td><td></td><td></td><td> 80</td>

<td>Tyr</td><td>Tyr</td><td>Cys</td><td>Gin</td><td>Gin</td><td>Tyr</td><td>Gly</td><td>How much</td><td>Cheese</td><td>Pro</td><td>Phe</td><td>Thr</td><td>Phe</td><td>Gly</td><td>Pro</td><td>Giy</td>

<td></td><td></td><td></td><td></td><td> 85</td><td></td><td></td><td></td><td></td><td> 90</td><td></td><td></td><td></td><td></td><td> 95</td><td></td>

<td>Thr</td><td>Lys</td><td>Val</td><td>Asp</td><td>How much</td><td>Lys</td><td>Arg</td><td>Thr</td><td>Val</td><td>Ala</td><td>Ala</td><td>Pro</td><td>Cheese</td><td>val</td><td>Phe</td><td>How much</td>

<td></td><td></td><td></td><td> 100</td><td></td><td></td><td></td><td></td><td> 105</td><td></td><td></td><td></td><td></td><td> 110</td><td></td><td></td>

<td>Phe</td><td>Pro</td><td>Pro</td><td>Cheese</td><td>Asp</td><td>Glu</td><td>Gin</td><td>Leu</td><td>, .ys</td><td>Cheese</td><td>dy</td><td>Thr</td><td>Ala</td><td>Cheese</td><td>Val</td><td>Val</td>

<td></td><td></td><td> 115</td><td></td><td></td><td></td><td></td><td> 120</td><td></td><td></td><td></td><td></td><td> 125</td><td></td><td></td><td></td>

<td>C ys</td><td>Leu</td><td>Leu</td><td>Asn</td><td>Asn</td><td>Phe</td><td>Tyr</td><td>Pro</td><td>Arg</td><td>Glu</td><td>Ais</td><td>Lys</td><td>Val</td><td>Gin</td><td></td><td></td>

<td></td><td> 130</td><td></td><td></td><td></td><td></td><td> 135</td><td></td><td></td><td></td><td></td><td> 140</td><td></td><td></td><td></td><td></td>

102

PL 214 003 Β1 &lt;210h 92 &lt;211&gt; 142 &lt;212&gt; PRT &lt;213&gt; Homo sapiens &lt;400&gt; 92

<td>Cheese 1</td><td>Pro</td><td>Gly</td><td>Thr</td><td>Leu 5</td><td>Cheese</td><td colspan="2">Leu Ser</td><td colspan="2">Pro Gly 10</td><td colspan="2">Glu Arg</td><td>Ala</td><td>Thr</td><td>Leu 15</td><td>Cheese</td>

<td>Cys</td><td>Arg</td><td>Ala</td><td>Cheese</td><td>Gin</td><td>Cheese</td><td>How much</td><td>Cheese</td><td>Cheese</td><td>Asn</td><td>Phe</td><td>Leu</td><td>Ala</td><td>Trp</td><td>Tyr</td><td>Gin</td>

<td></td><td></td><td></td><td> 20</td><td></td><td></td><td></td><td></td><td> 25</td><td></td><td></td><td></td><td></td><td> 30</td><td></td><td></td>

<td>Gin</td><td>Lys</td><td>Pro</td><td>Gly</td><td>Gin</td><td>Ala</td><td>Pro</td><td>Arg</td><td>Leu</td><td>Leu</td><td>How much</td><td>Tyr</td><td>Arg</td><td>Pro</td><td>Cheese</td><td>Cheese</td>

<td></td><td></td><td> 35</td><td></td><td></td><td></td><td></td><td> 40</td><td></td><td></td><td></td><td></td><td> 45</td><td></td><td></td><td></td>

<td>Arg</td><td>Ala</td><td>Thr</td><td>Gly</td><td>How much</td><td>Pro</td><td>Asp</td><td>Cheese</td><td>Phe</td><td>Cheese</td><td>Gly</td><td>Cheese</td><td>Gly</td><td>Cheese</td><td>Gly</td><td>Thr</td>

<td></td><td> 50</td><td></td><td></td><td></td><td></td><td> 55</td><td></td><td></td><td></td><td></td><td> 60</td><td></td><td></td><td></td><td></td>

<td>Asp</td><td>Phe</td><td>Thr</td><td>Leu</td><td>Thr</td><td>How much</td><td>Cheese</td><td>Arg</td><td>Leu</td><td>Glu</td><td>Pro</td><td>Glu</td><td>Asp</td><td>Phe</td><td>Al a</td><td>Leu</td>

<td> 65</td><td></td><td></td><td></td><td></td><td> 70</td><td></td><td></td><td></td><td></td><td> 75</td><td></td><td></td><td></td><td></td><td> 80</td>

<td>Tyr</td><td>Tyr</td><td>Cys</td><td>Gin</td><td>G1 n</td><td>Tyr</td><td>Gly</td><td>Thr</td><td>Cheese</td><td>Pro</td><td>Phe</td><td>Thr</td><td>Phe</td><td>Giy</td><td>Pro</td><td>Gly</td>

<td></td><td></td><td></td><td></td><td> 85</td><td></td><td></td><td></td><td></td><td> 90</td><td></td><td></td><td></td><td></td><td> 95</td><td></td>

<td>Thr</td><td>Lys</td><td>val</td><td>Asp</td><td>How much</td><td>Lys</td><td>Arg</td><td>Thr</td><td>Val</td><td>Ala</td><td>Ala</td><td>Pro</td><td>Cheese</td><td>Val</td><td>Phe</td><td>How much</td>

<td></td><td></td><td></td><td> 100</td><td></td><td></td><td></td><td></td><td> 105</td><td></td><td></td><td></td><td></td><td> 110</td><td></td><td></td>

<td>Phe</td><td>Pro</td><td>Pro</td><td>Cheese</td><td>Asp</td><td>Glu</td><td>Gin</td><td>Leu</td><td>Lys</td><td>Cheese</td><td>Gly</td><td>Thr</td><td>Ala</td><td>Cheese</td><td>Val</td><td>Val</td>

<td></td><td></td><td> 115</td><td></td><td></td><td></td><td></td><td> 120</td><td></td><td></td><td></td><td></td><td> 125</td><td></td><td></td><td></td>

<td>Cys</td><td>Leu</td><td>Wow</td><td>Asn</td><td>Asn</td><td>Phe</td><td>Tyr</td><td>Pro</td><td>Arg</td><td>Glu</td><td>Ala</td><td>Lys</td><td>Val</td><td>Gin</td><td></td><td></td>

130 135 140 &lt;210 93 &lt;211&gt; 146 &lt;212&gt; PRT &lt;213&gt; Homo sapiens &lt;400&gt; 93

<td>Gin 1</td><td>Cheese</td><td>Pro</td><td>Gly</td><td>Thr 5</td><td>Leu</td><td>Cheese</td><td>Leu</td><td>Cheese</td><td>Pro 10</td><td>Gly</td><td>Glu</td><td>Arg</td><td>Ala</td><td>Thr 15</td><td>Leu</td>

<td>Cheese</td><td>cys</td><td>Arg</td><td>Ala</td><td>Cheese</td><td>Gin</td><td>Cheese</td><td>Val</td><td>Cheese</td><td>Cheese</td><td>Tyr</td><td>Leu</td><td>Ala</td><td>Trp</td><td>Tyr</td><td>Gin</td>

<td></td><td></td><td></td><td> 20</td><td></td><td></td><td></td><td></td><td> 25</td><td></td><td></td><td></td><td></td><td> 30</td><td></td><td></td>

<td>Gin</td><td>Lys</td><td>Pro</td><td>Gly</td><td>Gin</td><td>Ala</td><td>Pro</td><td>Arg</td><td>Leu</td><td>Leu</td><td>How much</td><td>Tyr</td><td>Gly</td><td>Ala</td><td>Cheese</td><td>Cheese</td>

<td></td><td></td><td> 35</td><td></td><td></td><td></td><td></td><td> 40</td><td></td><td></td><td></td><td></td><td> 45</td><td></td><td></td><td></td>

<td>Arg</td><td>Ala</td><td>Thr</td><td>Gly</td><td>How much</td><td>Pro</td><td>Asp</td><td>Arg</td><td>Phe</td><td>Cheese</td><td>Gly</td><td>Cheese</td><td>Gly</td><td>Cheese</td><td>Gly</td><td>Thr</td>

<td></td><td> 50</td><td></td><td></td><td></td><td></td><td> 55</td><td></td><td></td><td></td><td></td><td> 60</td><td></td><td></td><td></td><td></td>

<td>Asp</td><td>Phe</td><td>Thr</td><td>Leu</td><td>Thr</td><td>How much</td><td>Cheese</td><td>Arg</td><td>Leu</td><td>Glu</td><td>Pro</td><td>Glu</td><td>Asp</td><td>Phe</td><td>Ala</td><td>Val</td>

<td> 65</td><td></td><td></td><td></td><td></td><td> 70</td><td></td><td></td><td></td><td></td><td> 75</td><td></td><td></td><td></td><td></td><td> 80</td>

Tyr Tyr Cys Gin Gin Tyr Gly Arg Ser Pro Phe Thr Phe Gly Pro Gly

PL 214 003 Β1

103

<td></td><td></td><td></td><td></td><td> 85</td><td></td><td></td><td></td><td></td><td> 90</td><td></td><td></td><td></td><td></td><td> 95</td><td></td>

<td>Thr</td><td>Lys</td><td>Val</td><td>ASP</td><td>how much</td><td>Lys</td><td>Arg</td><td>Thr</td><td>Val</td><td>Ala</td><td>Ala</td><td>Pro</td><td>Cheese</td><td>Val</td><td>Phe</td><td>How much</td>

<td></td><td></td><td></td><td> 100</td><td></td><td></td><td></td><td></td><td> 105</td><td></td><td></td><td></td><td></td><td> 110</td><td></td><td></td>

<td>Phe</td><td>Pro</td><td>Pro</td><td>Cheese</td><td>Asp</td><td>Glu</td><td>Gin</td><td>Leu</td><td>Lys</td><td>Cheese</td><td>Gly</td><td>Thr</td><td>Ala</td><td>Cheese</td><td>Val</td><td>Val</td>

<td></td><td></td><td> 1'15</td><td></td><td></td><td></td><td></td><td> 120</td><td></td><td></td><td></td><td></td><td> 125</td><td></td><td></td><td></td>

<td>Cys</td><td>Leu</td><td>Leu</td><td>Asn</td><td>Asn</td><td>Phe</td><td>Tyr</td><td>Pro</td><td>Arg</td><td>Glu</td><td>Ala</td><td>Lys</td><td>Val</td><td>Gin</td><td>Trp</td><td>Lys</td>