CONTRACEPTIVE VACCINE

Abstract: The instant invention is drawn to a sperm surface protein in substantially pure form selected from a human PH30 beta chain protein and a mouse PH30 beta chain proteins. Such proteins are useful as contraceptive vaccines in humans and mice respectively, and for identifying small molecules that will disrupt sperm-egg interaction and fertilization.

Description: TITLE OF THE INVENTION CONTRACEPTIVE VACCINE FIELD OF THE INVENTION The present invention provides sperm surface proteins and DNA sequences encoding the proteins which are useful in the prevention of fertilization. More particularly, the cloning and characterization of the mouse and human PH31 »eta chain genes, as well as their use as contraceptive vaccines, are described. BACKGROUND OF THE INVENTION Four methods of family planning are currently available in the U.S., sterilization, abstinence, abortion and contraception. Of these four birth control methods, contraception is the most widely utilized. Despite the substantial U.S. and global demand for contraception, the presently available methodologies fall short of market needs. Oral contraceptives and barrier methods dominate today's contraceptive market but have significant shortcomings. Oral contraceptives, though efficacious, are documented to be associated with significant side effects including increased risks of cardiovascular disease and breast cancer and are not recommended for women over the age of 35. Barrier methods, while safe, have failure rates approaching 20%. There is a clear need for increased availability of and improvements in contraceptives that offer superior safety, efficacy, convenience, acceptability and are affordable to women and men worldwide. Identification of novel approaches for controlling fertility is therefore necessary. Immunization of male and female animals with extracts of whole sperm is known to cause infertility. [Tung, K., et al., J. Reproductive Immunol., 1 ; 145-158 (1979); Menge, A., et al, Biol. of Reproduction, 20, 931-937 (1979)]. Moreover, men and women who spontaneously produce antisperm antibodies are infertile, but otherwise healthy. [Bronson, R., et al., Fert. and Sterile, 42, 171-183 (1984)]. Although the critical sperm antigens are unknown, these observations have led to the proposal that sperm proteins might be useful in the development of a contraceptives vaccine. In mammalian species, sperm proteins are believed to have a role in sperm adhesion to the zona pellucida of the egg. The PH30 protein is known to be involved in sperm egg binding and antibodies that bind to PH30 inhibit this interaction. PH30 is an integral membrane protein present on posterior head of sperm which mediates sperm-oocyte fusion. The PH30 protein consists of two immunologically distinct alpha and beta subunits. Both subunits are made as larger precursors and then finally processed in epididymis where sperm become fertilization competent. [Primakoff, P., et al.,J. Cell Biology, 104, 141 -149 (1987); Blobel, C.P., et al., J. Cell Biology, 1 1 1, 69-78 (1990)]. Monoclonal antibodies that recognize PH30 inhibit sperm-oocyte fusion in vitro, indicating its importance in fertilization [Primakoff, P., et al., /. Cell Biology, 104, 141 -149 (1987)]. Guinea pig PH30 alpha and beta chains have been cloned by Blobel et al. Mature PH30 alpha chain consists of 289 amino acids and encodes a transmembrane domain as well as an integral fusion peptide (82-102) that is similar to a potential fusion peptide of E2 glycoprotein of rubella virus. Guinea Pig PH30 beta chain has an open reading frame of 353 amino acids and also encodes a transmembrane domain. [Blobel C.P., et al., Nature, 356, 248-251 (1992)]. The predicted amino acid sequence of the PH30 beta chain protein contains significant homology to a class of proteins called disintigrins found in snake venom. These proteins .are known to bind to a family of proteins called integrins and prevent their normal functioning in cell adhesion (a well studied example is platelet aggregation). The N-terminal ninety amino acids integrin binding disintigrin domain of PH30 beta has been postulated to mediate the binding of PH30 to its putative integrin receptor on oocytes. The cloning and sequence determination of the mouse and human PH30 beta chain genes would permit novel approaches to the control of sperm egg binding and fusions. These approaches include, but are not limited to, eliciting an immune response directed at all or part of the PH30 beta chain protein and using the PH30 beta chain protein as part of a screen to identify small molecules that alter sperm egg interactions. Mammalian fertilization is, in most cases, species specific. Thus, the identification and isolation of sperm surface proteins essential for fertilization in species other than guinea pig would be useful for providing effective long lasting contraception in those species. Thus far, the lack of biochemical identification, isolation and cloning of candidate adhesion proteins of sperm has hindered scientists in developing effective contraceptives for humans as well as other mammalian species. SUMMARY OF THE INVENTION The instant invention relates to a sperm protein in substantially pure form selected from a human PH30 beta chain protein, a mouse PH30 beta chain protein or an amino acid sequence substantially homologous to either the human or mouse PH30 beta chain protein. In one embodiment of the invention is the sperm protein having an integrin binding sequence which is not TDE. In one class is the sperm protein wherein the integrin binding sequence is selected from FEE or QDE. In a subclass is the sperm protein which is the human PH30 beta chain protein. Illustrative of this subclass is the sperm protein having an integrin binding sequence that is FEE. Further illustrating the invention is a DNA sequence which encodes the sperm protein or a portion of the sperm protein sufficient to constitute at least one epitope. An illustration is the DNA sequence wherein the epitope is on the native protein. Exemplifying the invention is the DNA sequence which encodes all or a portion of human PH30 beta chain protein. An example of the invention is the DNA sequence, wherein the DNA encoding all or a portion of the human PH30 beta protein is characterized by the ability to hybridize, under standard conditions, to the DNA sequence shown in SEQ ID NO: 1. More particularly illustrating the invention is a contraceptive composition comprising a therapeutical ly effective amount of the protein, or a polypeptide having the substantially same amino acid sequence as a segment of the protein provided that the polypeptide is sufficient to constitute at least one epitope, and a pharmaceutically acceptable carrier. Another illustration is the contraceptive composition wherein the epitope is on the native protein. Further exemplifying the invention is the contraceptive composition, wherein the protein is the human PH30 beta chain protein. More specifically illustrating the invention is the contraceptive composition, wherein the protein is produced by expressing the gene encoding an immunogenic epitope of the sperm protein in a recombinant DNA expression vector. Specifically exemplifying the invention is a vector comprising an inserted DNA sequence encoding for the protein. A further illustration of the invention is the vector, wherein the inserted DNA sequence is characterized by the ability to hybridize, under standard conditions, to a DNA sequence selected from the DNA sequences of SEQ ID NO: 1 or SEQ ID NO: 3. Another example of the invention is a host that is compatible with and contains the vector. More specifically exemplifying the invention is a method of producing a human or mouse PH30 beta chain sperm protein, comprising the steps of culturing cells containing PH30 beta chain DNA and recovering the sperm protein from the cell culture. A further example is the method wherein the DNA encoding all or a portion of the PH30 beta chain protein is characterized by the ability to hybridize, under standard conditions, to a DNA sequence selected from the DNA sequences of SEQ ED NO: 1 or SEQ ID NO: 3. A more specific illustration is a method of contraception in a human or mouse subject in need thereof, comprising administering to the subject an amount of the sperm protein which is effective for the stimulation of antibodies which bind to the sperm protein in vivo, thereby preventing or substantially reducing the rate of sperm-egg fusion. Further illustrating the invention is the method wherein the sperm protein has an integrin binding sequence which is not TDE. Another illustration is the PH30 beta chain protein made by the process described. Another example is a DNA sequence as shown in Seq. ID No. 1 encoding human PH30 beta chain protein. Still further illustrating the invention is a purified anu isolated DNA sequence consisting essentially of a DNA sequence encoding a polypeptide having an amino acid sequence sufficiently duplicative of that of human or mouse PH30 beta to allow the possession of the biological property of initiating sperm-egg binding or promoting sperm-egg fusion. This biological activity can be determined using the in vitro sperm-oocyte binding/fusion assays [Primakoff, P., et al., J. Cell. Biol., 104: 141-149 (1987)]. More particularly exemplifying the invention is the DNA sequence wherein the amino acid sequence contains an integrin binding sequence which is not TDE. BRIEF DESCRIPTION OF THE DRAWINGS Figure 1 is a diagram representing the human PH30 beta cDNA gene sequence encoding the human PH-30 beta protein, and the deduced amino acid sequence of the human PH-30 beta protein present in three letter code. The sequence disclosure of Figure 1 is represented as SEQ ID NO: 1 and 2. Figure 2 is a diagram representing the mouse PH30 beta cDNA gene sequence, and the deduced amino acid sequence of the mouse PH-30 beta protein present in three letter code. The sequence disclosure of Figure 2 is represented as SEQ ID NO: 3 and 4. Figure 3 is a restriction MAP of the human PH30 beta cDNA sequence. Figure 4 is a restriction MAP of the mouse PH30 beta cDNA sequence. DETAILED DESCRIPTION OF THE INVENTION The subject invention relates to sperm surface proteins which are essential for fertilization, or portions thereof, and their use in contraceptive methods. A sperm surface protein is essential for fertilization if, for example, a monoclonal antibody to the protein or a polyclonal antibody raised against the purified protein, when bound to sperm, inhibits in vitro or in vivo fertilization or any step of in vitro fertilization. The process of fertilization is defined as the binding or fusion of two gametes (sperm and egg) followed by the fusion of their nuclei to form the genome of a new organism. The surface protein can be located in the plasma membrane of sperm and/or the inner acrosomal membrane. It can be a protein or glycoprotein. The isolated surface protein used for immunization can comprise the entire surface protein or some portion of the protein (external to the cell) which is immunogenic. Two such sperm surface proteins are the mouse and human PH30 beta chain sperm surface proteins. The PH30 beta genes encode proteins which are present on the surface of sperm cells and are essential for fertilization. As used herein, a protein or peptide is "substantially pure" when that protein or peptide has been purified to the extent that it is essentially free of other molecules with which it is associated in nature. The term "substantially pure" is used relative to proteins or pep- es with which the peptides of the instant invention are associated in natui , and are not intended to exclude compositions in which the peptide of the invention is admixed with nonproteinous pharmaceutical carriers or vehicles. As used herein, an amino acid sequence substantially homologous to a referent PH-30 beta protein will have at least 70% sequence homology, preferably 80%, and most preferably 90% sequence homology with the amino acid sequence of a referent PH-30 beta protein or a peptide thereof. For example, an amino acid sequence is substantially homologous to mouse PH-30 beta protein if, when aligned with mouse PH- 30 beta protein, at least 70% of its amino acid residues are the same. In addition, it is preferable that the substantially homologous amino acid sequence contains the integrin binding sequence. As used herein, a DNA sequence substantially homologous to a referent PH-30 beta protein will have at least 70%, preferably 80%, and most preferably 90% sequence homology with the DNA sequence of a referent PH-30 beta. Moreover, a DNA sequence substantially homologous to a referent PH-30 beta protein is characterized by the ability to hybridize to the DNA sequence of a referent PH30 beta under standard conditions. Standard hybridization conditions are described in Maniatis, T., et al. (1989) Molecular Cloning, Cold Spring Harbor Laboratory, Cold Spring Harbor, New York. An "expression vector" or "vector," as used herein, refers to a plasmid, bacteriophage, virus, or other molecule into which a gene of interest may be cloned, such that the appropriate signals for expression of that gene are present on that vector. The term "epitope," as used herein, refers to the minimum amount of PH30 beta sequence capable of producing an efficatious, i.e., contraceptive, immune response. The term "therapeutically effective amount," as used herein, means that amount of a drug or pharmaceutical agent that will elicit the biological or medical response that is being sought by a researcher or clinician. Production and Purification of Immunogen A preferred method for producing sperm surface proteins for use as a contraceptive immunogen is by recombinant DNA technology. To produce the protein using this technology it is necessary to isolate and clone DNA encoding the protein, or an immunogenic portion thereof. Those skilled in the art are familiar with a variety of approaches which can be used in an effort to clone a gene of interest. However, having nothing more than the isolated protein of interest, success in such an effort cannot be predicted with a reasonable degree of certainty. In the Examples which follow, Applicants describe the cloning and characterization of the mouse and human PH30 beta chain genes. The mouse and human PH30 beta chain genes were isolated using a cDNA encoding the guinea pig PH30 beta chain gene. The instant invention provides specific sequence information to permit targeted intervention in controlling fertility through anti PH30 directed immune responses inhibition of sperm-egg binding and triggering of post binding signaling and effective events. These sequences permit the generation of reagents for the isolation of oocyte proteins involved in sperm-egg interaction. The information presented in the Examples enable one skilled in the art to isolate and clone the mouse or human PH30 beta chain gene. For example, a cDNA library is prepared from testis or spermatogenic cells isolated from the mammal of interest (e.g., mouse, human). Such a cDNA library is then screened using, for example, labeled guinea pig PH30 DNA probes. DNA encoding all or a portion of human or mouse PH30 is characterized by the ability to hybridize to such a probe sequence under hybridization conditions such as those described in Example 1. Methods of labeling and screening by hybridization are well known in the art. Positive clones are analyzed, and a full length cDNA is constructed by conventional methods. The cloned gene, or portions thereof which encode an immunogenic region of the PH30 protein, can be expressed by inserting the coding region into an expression vector to produce an expression construct. Many such expression vectors are known to those skilled in the art. These vectors contain a promoter for the gene of interest as well as additional transcriptional and translational signals. Expression vectors for both eukaryotic host cells and prokaryotic host cells are widely available. The DNA expression construct is used to transform an appropriate host cell. Eukaryotic, in particular mammalian, host cells are often utilized for the expression of eukaryotic proteins. It has been found, for example, that eukaryotic proteins may exhibit folding problems when expressed in prokaryotic cells. In addition, production of authentic, biologically active eukaryotic proteins from cloned DNA sometimes requires post-translational modification such as disulfide bond formation, glycosylation, phosphorylation or specific proteolytic cleavage processes that are not performed in bacterial cells. This is especially true with membrane proteins. The sperm surface protein is produced using the transcriptional and translational components of the host cell. After an appropriate growth and expression period, the host cell culture is lysed and the sperm surface protein is purified from the lysate. Lysis buffers typically include non-ionic detergent, protease inhibitors, etc. From the solubilized cell extract, the sperm surface protein can be purified and isolated by physical and biochemical methods such as ultracentrifugation, column chromatography, high performance liquid chromatography, electrophoresis, etc. Alternatively, the sperm surface protein can be isolated by affinity chromatography using monoclonal or polyclonal antibodies [see Primakoff et al., Biol. of Reprod. 38, 921-934 (1988)]. Such methods for purifying proteins are well known to those skilled in the art. As mentioned above, antigenic portions or epitopes of the sperm surface protein are useful as immunogen, in addition to the full length protein. Antigenic fragments can be produced, for example, by proteolytic digestion of the full length protein, followed by isolation of the desired fragment. Alternatively, chemical synthesis can be used to generate the desired fragment starting with monomer amino acid residues. With respect to the PH30 protein, certain antigenic domains are preferred candidates for use in a contraceptive vaccine. As is discussed in greater detail in the Exemplification section which follows, the PH30 β subunit contains a domain which is highly conserved when compared to a class of proteins known as disintegrins. A peptide (or portion thereof) which is identical or substantially identical to this domain is preferred for use in the contraceptive methods of this invention. Substantially identical, as used in the preceding sentence, means that at least 70% of the amino acid sequence of the peptide is identical to the corresponding portion of the PH30 β disintegrin domain. Disintegrins are found in snake venom, for example, and are known to bind to a class of platelet surface proteins known as integrins. The binding of disintegrins to integrins has been shown to inhibit blood clotting. By analogy, peptides corresponding to the PH30 β disintegrin domain are predicted to be active in sperm-egg binding and fusion. Contraceptive Vaccine Once the sperm surface protein has been produced and purified, a vaccine can be produced by combining the sperm surface protein or portion thereof with a suitable carrier for administration to a subject for immunization. For successful vaccine development it is necessary that the immunogen exhibit tissue specificity, that is, it is expressed on the target tissue only and must be essential for the process of reproduction. It is known that the PH30 protein, which is expressed only on sperm, is involved in sperm egg binding and antibodies that bind to PH30 inhibit that interaction. The cloning and characterization of human PH30 beta permits novel approaches for using PH30 as a target to control human fertility. PH30 beta protein or peptides can be used directly as an antigen to elicit an immune response directed to the whole or a relevant part of the PH30 beta chain protein. Testing of these approaches requires availability of sufficient quantities of PH30 beta protein. The cloning and sequencing of the mouse and human PH30 beta chain provides information necessary to recombinantly express all or part of the PH30 beta protein. These expressed proteins are used with or without adjuvant to immunize women or female mice. The elicited humoral immune responses are monitored by assays that use PH30 beta as antigen. Secreted antibodies in the female reproductive system will bind to the sperm head and disrupt fertilization. The availability of the recombinant mouse PH30 beta protein permits establishment of an animal model system for testing efficacy, reversibility and safety of specific methods of controlling fertility based on PH30. A vaccine can contain one or more sperm surface proteins. Sperm surface proteins of the present invention can be combined with adjuvants which contain non-specific stimulators of the immune system. Proper use of adjuvants can induce a strong antibody response to foreign antigens (i.e., sperm surface proteins). The action of adjuvants is not fully understood, but most adjuvants incorporate two components. One is a substance designed to form a deposit which protects the antigen from catabolism. Two methods of forming a deposit are to use mineral oils or aluminum hydroxide precipitates. With mineral oils, such as Freund's adjuvant, the immunogen is prepared in a water-in-oil emulsion. For aluminum hydroxide, the immunogen is either adsorbed to preformed precipitants or is trapped during precipitation. The second component required for an effective adjuvant is a substance that will stimulate the immune system nonspecifically. These substances stimulate the production of a large set of soluble peptide factors known as lymphokines. In turn, lymphokines stimulate the activity of antigen-processing cells directly and cause a local inflammatory reaction at the site of injection. A component of lipopolysaccharide known as lipid A is commonly used. Lipid A is available in a number of synthetic and natural forms that are much less toxic than lipopolysaccharides, but still retain most of the desirable adjuvant properties of the lipopolysaccharide molecules. Lipid A compounds are often delivered using liposomes. The two bacteria that are commonly used in adjuvants as non-specific stimulants are Bordatella pertussis and Mycobacterium tuberculosis. When used as whole bacteria, they must be heat-killed prior to use. The immunomodulatory mediators of B. pertussis include a lipopolysaccharide component and the pertussis toxin. The pertussis toxin has been purified and is available commercially. M. tuberculosis is commonly found in complete Freund's adjuvant. The most active component of M. tuberculosis has been localized to muramyl dipeptide which is available in a number of forms. Immunizations (Inoculation and Booster Shots) The subject to be immunized can be any mammal which possesses a competent immune system. Examples of subject mammals include humans and domestic animals (e.g. dogs, cats, cows, horses, etc.), as well as animals intended for experimental or other purposes (e.g., mice, rats, rabbits, etc.). Two different criteria are important to consider in determining the proper dose for the initial immunization. First, the optimum dose to achieve the strongest response and second, the minimum dose likely to induce the production of useful polyclonal antibodies. Much of the injected material will be catabolized and cleared before reaching the appropriate target immune cell. The efficiency of this process will vary with host factors, the route of injection, the use of adjuvants, and the intrinsic nature of the surface protein injected. Thus, the effective dose delivered to the immune system may bear little relationship to the introduced dose and consequently dose requirements must be determined empirically. These determinations can be readily made by one skilled in the art. Secondary injections and later boost can be given with amounts similar to or less than the primary injection. The route of injection is guided by three practical decisions: 1) what volume must be delivered; 2) what buffers and other components will be injected with the immunogen; and 3) how quickly should the immunogen be released into the lymphatics or circulation. For example, with rabbits, large volume injections normally are given at multiple subcutaneous sites. For mice, large volumes are only possible with intraperitoneal injections. If adjuvants or paniculate matter are included in the injection, the immunogen should not be delivered intravenously. If a slow release or the inoculant is desired, the injections should be done either intramuscularly or intradermally. For immediate release, use intravenous injections. Primary antibody responses often are very weak, particularly for readily catabolized, soluble antigens. Hence, secondary or booster injections are required after the initial immunization. A delay is needed before reintroducing the protein into a primed subject. A minimum of 2 or 3 weeks is recommended but greater intervals are possible. The antibody responses to secondary and subsequent injections is much stronger. Higher titers of antibody are reached, but more importantly, the nature and quantity of the antibodies present in serum changes. These changes yield high-affinity antibodies. The intervals between secondary, tertiary and subsequent injections may also be varied, but usually need to be extended to allow the circulating level of antibody to drop enough to prevent rapid clearance of newly injected antigen. Subsequent booster injections will be required to increase reduced circulating antibody for continued contraception. The actual intervals for these injections will differ from species to species. However, the intervals can be determined by one skilled in the art by monitoring serum levels of sperm surface protein antibodies. In another embodiment, subjects can be administered with alloantisera, or monoclonal antibodies, directed to a sperm surface protein to achieve contraception. The alloantiserum is raised in another individual of the same species, isolated from the serum of the individual and prepared in a suitable carrier for injection into the recipient subject. Those skilled in the art are familiar with methods for preparing and formulating monoclonal antibodies for administration. There is convincing evidence that naturally occurring antibodies to sperm cause infertility in women [Bronson, R.A., et al., Fertility and Sterility, 42: 171-183 (1984)]. This infertility is better correlated with the antibody titers in cervical mucus than with the serum [Clark, G.N., Amer. J Reprod. Immunol, 5: 179- 181 (1984)]. Presence of anti-sperm antibodies in the cervical mucus of infertile women results in poor sperm penetration through the cervical mucus and agglutination of the sperm, thereby reducing the number of sperm available for fertilization. Thus, success of a contraceptive vaccine depends in particular on the generation of mucosal immune responses involving sustained titers of anti- sperm antibodies in the female reproductive tract. Generally, local application of the antigen is an effective way to stimulate an antibody response by that mucosa [Mestecky, J., J Clin. Immunol, 7: 265-276 (1987)]. However, local mucosal immunization is ineffective in female reproductive tract due to the barrier function of the luminal epithelium and to rapid loss of antigen from the lumen of reproductive tract. Stability and adhesiveness of the antigen on the mucosal surface is important for the induction of the mucosal immune responses [de Aizpurua, H.J. and Russell-Jones, G.J., J Exp. Med., 167: 440 (1988)]. Adhesive antigens are critical to successful mucosal immunization, not only because they are effective mucosal immunogens themselves, but also because they are carrier proteins for other antigens. Cholera toxin is a potent immunogen when given mucosally, but acts as an adjuvant when given in combination with other antigens [McKenzie, S.J. and Halsey, J.A., J. Immunol, 133: 1818 (1984)]. Effective immunization is also dependent on the stability of the antigen on a mucosal surface. Many antigens for use in mucosal vaccines are poorly immunogenic because they are unable to survive in the acidic and proteolytic conditions of the mucosal surface [O'Hagen, D.T., Curr. Opin. Infect. Dis., 3:393 (1990)]. The DL-lactide- co-glycolide (DL-PLG) microsphere, microparticle carrier system is one of the most suitable systems for mucosal immunization. DL-PLG microspheres protect the antigen at mucosal surface and are taken up by the mucosal lymphoid tissues where they induce mucosal immunity [Eldridge, J.H. et al, Curr. Top. Microbiol. Immunol, 146: 59 (1989)]. Liposomes and inactivated micro-organisms also are used as microparticle carriers. Some parenteral adjuvants such as Avridine, a lipoidal amine and muramyl dipeptide (MDP), the active component of mycobacteria in Freund's complete adjuvant, also have been shown to be active as oral mucosal adjuvants and enhance mucosal immunization [Anderson, A.O. and Reynolds, J.A., J. Reticuloendothel Soc, 26(suppl): 667 (1979); Taubman, M.A., et al., Ann. NY Acad. Sci., 409: 637 (1983)]. Development of mucosal immune responses in female reproductive tract are optimized by using various adjuvants, micro particle carriers, by immunizing at local or remote mucosal surfaces or by combination of parenteral and mucosal immunization. Utility of PH30 beta in Identification of Small Molecules that will Disrupt Sperm-egg Interaction and Fertilization The comparison of the protein sequences of both mouse and human PH30 beta chain genes shows significant homology to a class of proteins called disintegrins found in the snake venoms. These proteins are known to bind a family of cell surface molecules called integrins and prevent their normal function in cell adhesion. On the basis of these homologies it is reasonable to conclude that the PH30 receptor on the oocyte is an integrin. Comparisons of the disintegrin domain sequences of guinea pig, mouse and human PH30 beta chain genes show significant differences in their putative ligand binding domain. In particular, the sequences in this region are different from other disintegrins and among the three species. The recombinant mouse and human PH30 beta proteins are used to make affinity resins to purify, identify and characterize mouse and human PH30 receptors. The recombinant PH30 beta also are used to determine its relative affinity to other integrins expressed in other tissues and are used as a ligand for cloning of the PH30 receptor. Since the integrin recognition sequences in PH30 beta are species specific, the sequence information is necessary to identify small molecules that disrupt fertilization in a species specific manner. The recombinant mouse and human PH30 beta are used to set up screens to identify small molecules that act either as antagonist to PH30 receptor and disrupt PH30 binding or act as an agonist and stimulate PH30 receptor inducing transmembrane signaling, egg cortical granule release and zona reaction thus making the egg impenetrable for fertilization. The present invention is further illustrated in the following exemplification. EXAMPLE 1 Isolation of DNA Encoding Mouse and Human PH30 beta A. cDNA Library Plating One million independent recombinant bacteriophage from both a human testis cDNA library in λgt 11 (Clontech, Palo Alto, CA.) and mouse testis cDNA library (Stratagene La Jolla, CA.) in UNI-ZAP XR were plated. Plaque lifts were done in duplicate by placing a nitrocellulose filter on the plate for two minutes, and treating the filter with denaturing solution (0.5M NaOH, 1.5M NaCl), neutralization buffer (0.5M Tris pH 7,5, 1.5M NaCl) and 2X SSC (3M NaCl, 0.35M sodium citrate pH 7.0) for two minutes each. The filters were dried for thirty minutes at room temperature and then baked for two hours at 80° C in a vacuum oven. B. Generation of Probe: A guinea pig PH30 beta cDNA was isolated by RT-PCR (reverse transcriptase-polymerase chain reaction) as a 1020 bp (base pairs), HindHI/Bam HI fragment, containing 94% of the coding sequence. This fragment was subcloned into pBluescript SK+ vector (Stratagene, La Jolla, CA) and verified by sequence analysis. A probe was made by nick translating the purified 1020 bp guinea pig PH30 beta fragment. The filters were probed at 42°C for fifteen hours in hybridization solution (7mM Tris pH 7.5, 40% formamide, 4X SSC, 0.8X Denhard's, 20 μg/ml of salmon sperm DNA and 10% Dextran sulfate) containing 1 θ6 cpm (counts per minute)/ml of the labeled probe. The filters were washed twice at room temperature for fifteen minutes each with 2X SSC/0.2% SDS (sodium dodecyl sulfate), then twice at room temperature with 0.2X SSC/0.1 %SDS, then once at 42°C for 30 minutes with 0.1X SSC/0.1%SDS. The filters were exposed to XAR film (Eastman Kodak Co, Rochester, NY) for 15 hours. The positive plaques were picked into 1 ml of SM ( 0.1M NaCl, lOmM Magnesium Sulphate, 2% gelatin, 50mM Tris pH 7.5) and screened again as described above. After four rounds of screening, the purified plaques were obtained. Purified plaques of mouse testicular library were subcloned into pBluescript SK+ vector using the EX ASSIT helper phage and SOLR cells (Stratagene, La Jolla, CA). DNA from the purified plaques of human testicular library was isolated using light PLG 2 tubes and following manufacturer's (Clontech, Palo Alto, CA) directions. The DNA was then digested with the restriction enzyme EcoRI and ligated into pBluescript SK+ and was used to transform competent E. coli strain HB101 cells. C. DNA Sequencing and Analysis: Cloned inserts were sequenced on both strands using the Sequenase kit (United States Biochemical, Cleveland, Ohio). Sequences were analyzed by searching GeneBank and EMBL DNA sequence database using the FASTA program (University of Wisconsin, Genetics Computer Group) and sequence comparisons were done using the GAP program. D. Characterization of cDNA Clones: The screening of the mouse testicular library with a 1020 bp guinea pig PH30 beta probe resulted in the isolation of a 1.7 kb (kilo base pair) cDNA clone. This cDNA clone contains a 1371 nucleotide open reading frame and a 329 nucleotide 3' untranslated region. When mature parts of the guinea pig and mouse PH30 beta were compared, the mouse PH30 beta clone showed a maximum of 63% identity to guinea pig PH30 beta at the nucleotide level. The amino terminal 103 residues of the deduced 457 amino acid sequence represents the precursor regions of the mouse PH30 beta that are cleaved off at sperm maturation. At the amino acid level the mature mouse, and guinea pig PH30 betas were 54% identical with all the cysteines lining up. The human testicular cDNA library screening identified a 2.331 kb cDNA which contains an open reading frame of 1959 nucleotides and 372 nucleotide 3' untranslated region. The human PH30 beta clone was 63 and 67% identical in its open reading frame to mouse and guinea pig PH30 beta genes, respectively. Comparison of the derived 653 amino acid sequence with the mouse and guinea pig PH30 beta indicates that the amino terminal 299 represents the precursor and carboxy terminal 354 amino acids represent the mature part of human PH30 beta respectively. The amino acid sequence of the mature human PH30 beta was 54% homologous to mature guinea pig and mouse PH30 beta proteins. Protein sequence comparison of mouse and human PH30 beta to guinea pig PH30 beta and snake venom disintegrins indicated significant homology. This analysis revealed similar structural organization and indicated the presence of metalloprotease and disintegrin domains in these proteins. Metalloprotease domains of mouse and human PH30 beta shared significant similarity with the metalloprotease domains of guinea pig PH30 beta but less similarity to the metalloprotease domain of guinea pig PH30 alpha or other disintegrins. The active site signature sequence of zinc-dependent metalloproteases is present in PH30 alpha and the snake venom disintegrins, Jararhagin and Trigramin. [Wolfsberg, T.G., et al., Proc. Natl. Acad. Sci. USA 90: 10783-10797 ( 1993)]. Similar to guinea pig PH30 beta, the mouse and human metalloprotease domain lacks the active site signature sequence and both were 80% identical to guinea pig PH30 beta and only 30% identical to guinea pig PH30 alpha metalloprotease active site sequence. Human and guinea pig PH30 beta metalloprotease domains were 60% identical. Similar to guinea pig PH30 beta, the mouse and human PH30 beta also contain a disintegrin domain. The disintegrin domain in mouse PH30 beta contains 91 amino acids (residues 11 1-202) and in human, 93 amino acids (residues 299-392). Most disintegrins of snake venom contain a consensus integrin binding sequence RGD. Another family of snake venom disintegrins that are linked to a carboxyl terminus cysteine rich domain, lack the RGD tripeptide but contain a unique tripeptide and adjacent cysteine. Guinea pig, mouse and human PH30 beta proteins also do not contain RGD tripeptide and share more similarity with this later family of disintegrins. These snake venom disintegrins and disintegrin domains of guinea pig, mouse and human PH30 beta contain a negatively charged residue at the carboxyl end of the tripeptide sequence. The integrin binding sequence of guinea pig PH30 beta is TDE. One skilled in the art would have expected that the integrin binding site of PH30 beta of other mammalian species would also be TDE. However, after isolation of human and mouse PH30 beta, it was found that this was not the case. It was unexpectedly discovered that the critical sequence at the integrin binding site was not conserved. Comparisons of guinea pig, mouse and human PH30 beta disintegrin domains showed significant variation in their putative integrin binding sequences although the carboxy terminus end of these domains were identical. The putative integrin binding residues in PH30 beta were QDE in mouse and FEE in human. These differences in the integrin binding sequences between species were an unexpected and surprising finding. Both mouse and human PH30 beta contain an epidermal growth factor like repeat and a transmembrane domain that are 60% identical to similar regions of guinea pig PH30 beta. EXAMPLE 2 Cloning of the 5' end of Mouse and Human PH30 Beta The 5' ends of mouse and human PH30 beta were cloned using the Gibco BRL "5' RACE System for Rapid Amplification of cDNA Ends" and following manufacturer's protocols. 2 oligonucleotides were synthesized for each template. Oligo 1 was an antisense primer and Oligo 2 was also an antisense primer, internal to oligo 1 , and contained in the CAU sequences on the 5' end to facilitate cloning. Oligo 1 was annealed to mouse or human testis mRNA and a cDNA copy was made using Superscript II Reverse Transcriptase. The mRNA template was degraded with Rnase H. The single strands cDNA copy was purified with GlassMAX Spin columns and was then tailed on the 3' end with dCTP and terminal transferase. The tailed cDNA copy was then amplified using a supplied anchor primer that contains the 5'CAU cloning site and oligo 2. The amplification system was Taq polymerase. The amplified product was then gel purified, treated with Uracil DNA Glycosylase, subcloned into the vector pAMPl and then transformed into competent E. coli DH5 cells. Colonies were identified which had subcloned fragment and these colonies were sequenced as described previously. The complete mouse cDNA sequence and the deduced amino acid sequence of the mouse PH30 beta protein is shown in SEQ ID NO: 5 and SEQ ID NO: 6. The complete human cDNA sequence and the deduced amino acid sequence of the human PH30 beta protein is shown in SEQ ID NO: 7 and SEQ ID NO: 8. At the nucleotide level, the complete human PH30 beta shares 68% identity with mouse and 68.6% identity with guinea pig PH30 beta, respectively. Mouse and guinea pig DNA sequences are 65.5% identical. The amino acid sequence of the human PH30 beta is 58.9% identical to mouse and 56.5% identical to guinea pig PH30 beta. At the amino acid level, the mouse and guinea pig PH30 beta are 55.2% identical. EXAMPLE 3 Contraceptive Vaccination by the Administration of PH30 beta Protein Female or male mice (about 7 weeks old at the time of first injection) receive two injections of PH30 beta in the amounts stated below. Recombinant or native PH30 beta, purified from cell line or sperm by mAb-affinity chromatography or biochemical methods, shows at least 90% purity (i.e., no more than 10% detectable contaminants) using silver-staining of purified protein on SDS gels. Purity of each PH30 preparation used for immunization of females or males is verified by SDS polyacrylamide gel electrophoresis and silver staining. The affinity-purified PH30 beta, in 0.375 ml phosphate-buffered saline (PBS) containing 3 mM octyglucoside (OG) is emulsified with 0.375 ml complete Freund's adjuvant (CFA). Each animal receives 0.1 ml of the emulsion subcutaneously in the back and 0.05 ml intramuscularly in a rear leg. About 3 weeks later, the same amount of PH30 beta in PBS and 3 mM OG is emulsified with incomplete Freund's adjuvant (IFA), and is injected in the same sites in each animal. Control females and males receive the same injections on the same schedule and containing PBS and 3 mM OG and CFA or IFA, but lacking PH30 beta. To allow the injected females to mate, about 6 weeks after the initial injection they are housed with males for 10 days. Each cage contains one male (13 weeks old) , one PH30 beta immunized female, and from 2-4 control injected females. 24 hours after the grouping, females are checked visually daily for the vaginal plugs. Two weeks after the initiation of the mating the, females are removed into individual cages. After three weeks the pregnant females having litters and progeny are counted. To allow the injected males to mate, about six weeks after the initial injection, each injected male is housed with two females (10-13 weeks) for 10 days. The females and males are then separated and after an additional 3 weeks pups are counted. EXAMPLE 4 Use of PH30 Disintegrin Peptides as Inhibitor of Sperm Fusion to Egg Plasma Membrane Peptides from the PH30 β disintegrin domain are tested for inhibition of sperm binding to the egg plasma membrane. The fusion inhibition assay is carried out as follows. Young female mice (8-10 weeks of age) are injected with 5 units of pregnant mare's serum (PMS) in 0.9 NaCl intraperitoneally. 48 hours later, the mice are injected IP with 5 units of hCG ( human chorionic gonadotrophin) in 0.9% NaCl to trigger super ovulation. 14-16 hours after hCG injection, the ovulated oocytes are collected and treated with hyaluronidase to remove cumulus cells. The zona pellucida is removed with a mixture of proteases. The zona pellucida free eggs are incubated in culture media with peptide at a specified concentration for 30 minutes [Hogan, B.. et al., Manipulating The Mouse Embryo, 91 -101 , (1986)]. Sperm collected from the epididymis of male mice is capacitated by incubation and acrosome reacted as described by Fleming and Yanagimachi [Gamete Res. 4, 253-273 (1981)] and added to the eggs and incubated for 15 minutes. The eggs are then transferred to a sperm free culture medium and incubated for an additional 1 hour and 45 minutes. The eggs are then fixed and stained as described by Primakoff et al., [J. Cell Biol. 104, 141 (1987)]. The total number of swollen sperm heads are then counted. Swollen sperm heads are an indication that the sperm and egg have fused. On the basis of these observations, several indices are calculated. The fertilization index (F.I.) is determined by dividing the total number of swollen heads by the total number of eggs. The fertilization rate (F.R.) is the percentage of eggs fertilized. The percent inhibition is determined by dividing the fertilization index of the experimental peptide by the fertilization index of the control peptide. The PH30 β disintegrin domain represents an epitope which is critical in sperm-egg fusion. Antibodies which bind specifically to this epitope block sperm/egg fusion. EXAMPLE 5 Use of PH30 beta to Identify Small Molecules that will disrupt sperm-egg Interaction and Fertilization A. Identification of PH30 beta receptor antagonists: Identification of compounds that specifically interfere with the binding of PH30 to their receptor on the egg, has been limited due to unavailability of the sufficient quantities of PH30 protein and normal human eggs. The availability of the rPH30 beta facilitates the identification and cloning of PH30 beta receptor integrin cDNAs. These PH30 beta receptor cDNAs are used to generate recombinant PH30 beta receptors. The alternative source of PH30 beta receptors facilitates identification of substances that affect the binding of PH30 beta to its receptors. Using conventional methods, the Chinese Hamster Ovary cells are transfected with cDNAs encoding the PH30 beta receptor to produce a stable transformed cell which expresses human PH30 beta receptor integrin in large quantities. Such a transformed cell provides a consistent source of recombinant PH30 beta receptors and is useful in the characterization of the binding of PH30 beta to its receptor and for establishing assays to screen for compounds that inhibit PH30 binding to its receptor. Selectivity of the compounds to PH30 beta receptor is examined by using cell lines that express other integrin receptors that contain the same beta subunit and closely related alpha chain. Compounds that specifically inhibit PH30 beta/receptor interaction are tested further in biological assays, like inhibition of sperm-egg fusion assay and egg cortical granule release assay to determine their efficacy in inhibiting fertilization. B. Protocol for PH30 beta antagonist screen: Cells expressing PH30 beta receptor are treated with extraction buffer (50 mM Tris pH 7.6, 100 mM n-Octyl β-D- Glucopyranoside, 150 mM NaCl, 1 mM MgCl2 and 1 mM CaCl2) and soluble material is separated by centrifugation and stored frozen at -80 °C. In an assay tube the 15 μl water, 80 μl of assay buffer (125 mM Tris pH 7.6, 187.5 mM NaCl, 1.25 mM CaCl2, 1.25 mM MgCl2 and 1.25%BSA) and 5 μl of sample compound or control (40 μM of cold PH30 beta) are added and mixed with 50 μl of 125I_PH30 beta (final concentration 40 pM) and 50 μl of cell extract (final protein concentration 250 μg/ml). The tubes are incubated at room temperature for 1 hour. Following incubation the samples are harvested using Tomtec Mach II- 6x 16 cell harvester and printed filtermat cat. # 1205-404. Filters are dried and counted in LKB/Wallac Beta Plate counter. Calculations and Interpretations: % Inhibition = CPMavg total binding - CPMavg sample XI 00 CPMavg total binding - CPMavg positive control When % inhibition > 60 and the inhibition is dose related, the sample will be considered active. C. Sperm-Oocyte fusion assay: Young female mice (approximately 8-10 weeks of age) are injected with 5 units of pregnant mare's serum (PMS) in 0.9 NaCl intraperitoneally. 48 hours later, the mice are injected IP with 5 units of hCG (human chorionic gonadotrophin) in 0.9% NaCl to trigger super ovulation. 14-16 hours after hCG injection, the ovulated oocytes are collected and treated with hyaluronidase to remove cumulus cells. Zona pellucida is removed by treating eggs briefly with 0.1 mg/ml of chymotrypsin. Oocytes are washed with Hepes buffered culture medium and are loaded with a fluorescent stain 4',6-diamidino-2-phenylindole dihydrochloride (DAPI) by incubating at 37°C for 30 minutes. Oocytes are then washed with medium and incubated with rPH30 beta or inhibitor compound for 30 minutes followed by another 30 minute incubation with lxlθ4 sperms that have been previously capacitated by incubating with calcium ionophore. After incubation, the oocytes are washed, mounted and examined by light microscopy and scored for the presence of fluorescent swollen sperm heads with associated tails in cytoplasm. Fertilization rate = number of eggs fused XI 00 (results expressed as % number of eggs tested fertilization) In the absence of any inhibitor > 90% oocytes are fertilized. When the sperm-oocyte fusion is inhibited >60% and the inhibition is dose related the compound will be considered active. While the invention has been described and illustrated with reference to certain preferred embodiments thereof, those skilled in the art will appreciate that various changes, modifications and substitutions can be made therein without departing from the spirit and scope of the invention. It is intended, therefore, that the invention be limited only by the scope of the claims which follow and that such claims be interpreted as broadly as is reasonable. SEQUENCE LISTING ) GENERAL INFORMATION: (l) APPLICANT: ALVES, KENNETH GUPTA, SUNIL K. HOLLIS, GREGORY F. (ll) TITLE OF INVENTION: CONTRACEPTIVE VACCINE (ill) NUMBER OF SEQUENCES: 8 (IV) CORRESPONDENCE ADDRESS: (A) ADDRESSEE: MARY A. APPOLLINA (B) STREET: P.O. BOX 2000, 125 E. LINCOLN AVENUE (C) CITY: RAHWAY (D) STATE: NJ (E) COUNTRY: USA (F) ZIP: 07065 (v) COMPUTER READABLE FORM: (A) MEDIUM TYPE: Floppy disk (B) COMPUTER: IBM PC compatible (C) OPERATING SYSTEM: PC-DOS/MS-DOS (D) SOFTWARE: Patentin Release #1.0, Version #1.30 (vi) CURRENT APPLICATION DATA: (A) APPLICATION NUMBER: US (B) FILING DATE: (C) CLASSIFICATION: 'vi l ATTORNEY/AGENT INFORMATION: (A) NAME: APPOLLINA, MARY A iB) REGISTRATION NUMBER: 34,037 'O REFERENCE/DOCKET NUMBER: 192 4Y (IX) TELECOMMUNICATION INFORMATION: (A) TELEPHONE: (908)594-3462 (B) TELEFAX: (908)594-4720 INFORMATION FOR SEQ ID NO: 1: (l) SEQUENCE CHARACTERISTICS: (A) LENGTH: 2373 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: double (D) TOPOLOGY: linear (ll ) MOLECULE TYPE: cDNA (xi ■ SEQUENCE DESCRIPTION: SEQ ID NO: 1 : GGCCAAGATT TTCAGAATTT CTGCCACTAC CAAGGGTATA TTGAAGGTTA TCCAAAATCT 60 GTGGTGATGG TTAGCACATG TACTGGACTC AGGGGCGTAC TACAGTTTGA AAATGTTAGT 120 TATGGAATAG AACCCCTGGA GTCTTCAGTT GGCTTTGAAC ATGTAATTTA CCAAGTAAAA 180 CATAAGAAAG CAGATGTTTC CTTATATAAT GAGAAGGATA TTGAATCAAG AGATCTGTCC 240 TTTAAATTAC AAAGCGCAGA GCCACAGCAA GATTTTGCAA AGTATATAGA AATGCATGTT 300 ATAGTTGAAA AACAATTGTA TAATCATATG GGGTCTGATA CAACTGTTGT CGCTCAAAAA 360 GTTTTCCAGT TGATTGGATT GACGAATGCT ATTTTTGTTT CATTTAATAT TACAATTATT 420 CTGTCTTCAT TGGAGCTTTG GATAGATGAA AATAAAATTG CAACCACTGG AGAAGCTAAT 480 GAGTTATT .C ACACATTTTT AAGATGGAAA ACATCTTATC TTGTTTTACG TCCTCATGAT 540 GTGGCATTTT TACTTGTTTA CAGAGAAAAG TCAAATTATG TTGGTGCAAC CTTTCAAGGG 600 AAGATGTGTG ATGCAAACTA TGCAGGAGGT GTTGTTCTGC ACCCCAGAAC CATAAGTCTG 660 GAATCACTTG CAGTTATTTT AGCTCAATTA TTGAGCCTTA GTATGGGGAT CACTTATGAT 720 GACATTAACA AATGCCAGTG CTCAGGAGCT GTCTGCATTA TGAATCCAGA AGCAATTCAT 780 TTCAGTGGTG TGAAGATCTT TAGTAACTGC AGCTTCGAAG ACTTTGCACA TTTTATTTCA 840 AAGCAGAAGT CCCAGTGTCT TCACAATCAG CCTCGCTTAG ATCCTTTTTT CAAACAGCAA 900 GCAGTGT3TG GTAATGCAAA GCTGGAAGCA GGAGAGGAGT GTGACTGTGG GACTGAACAG 960 GATTGTGC C TTATTGGAGA AACATGCTGT GATATTGCCA CATGTAGATT TAAAGCCGGT 1020 TCAAACT3TG CTGAAGGACC ATGCTGCGAA AACTGTCTAT TTATGTCAAA AGAAA.GAATG 1080 TGTAGGCCTT CCTTTGAAGA ATGCGACCTC CCTGAATATT GCAATGGATC ATCTGCATCA 1140 TGCCCAGAAA ACCACTATGT TCAGACTGGG CATCCGTGTG GACTGAATCA ATGGATCTGT 1200 ATAGATGGAG TTTGTATGAG TGGGGATAAA CAATGTACAG ACACATTTGG CAAAGAAGTA 1260 GAGTTTGGCC CTTCAGAATG TTATTCTCAC CTTAATTCAA AGACTGATGT ATCTGGAAAC 1320 TGTGGTATAA GTGATTCAGG ATACACACAG TGTGAAGCTG ACAATCTGCA GTGCGGAAAA 1380 TTAATAPGTA AATATGTAGG TAAATTTTTA TTACAAATTC CAAGAGCCAC TATTATTTAT 1440 GCCAACATAA GTGGACATCT CTGCATTGCT GTGGAATTTG CCAGTGATCA TGCAGACAGC 1500 CAAAAGATGT GGATAAAAGA TGGAACTTCT TGTGGTTCAA ATAAGGTTTG CAGGAATCAA 1560 AGATGTGTGA GTTCTTCATA CTTGGGTTAT GATTGTACTA CTGACAAATG CAATGATAGA 1620 GGTGTATG A ATAACAAAAA GCACTGTCAC TGTAGTGCTT CATATTTACC TCCAGATTGC 1680 TCAGTTCAAT CAGATCTATG GCCTGGTGGG AGTATTGACA GTGGCAATTT TCCACCTGTA 1740 GCTATACC.AG CCAGACTCCC TGAAAGGCGC TACATTGAGA ACATTTACCA TTCCAAACCA 1800 ATGAGATGGC CATTTTTCTT ATTCATTCCT TTCTTTATTA TTTTCTGTGT ACTGATTGCT 1860 ATAATGGTGA AAGTTAATTT CCAAAGGAAA AAATGGAGAA CTGAGGACTA TTCAAGCGAT 1920 GAGCAACCTG AAAGTGAGAG TGAACCTAAA GGGTAGTCTG GACAACAGAG ATGCCATGAT 1980 ATCACTTCTT CTAGAGTAAT TATCTGTGAT GGATGGACAC AAAAAAATGG AAAGAAAAGA 2040 ATGTACATTA CCTGGTTTCC TGGGATTCAA ACCTGCATAT TGTGATTTTA ATTTGACCAG 2100 AAAATATGAT ATATATGTAT AATTTCACAG ATAATTTACT TATTTAAAAA TGCATGATAA 2160 TGAGTTTTAC ATTACAAATT TCTGTTTTTT TAAAGTTATC TTACGCTATT TCTGTTGGTT 2220 AGTAGACACT AATTCTGTCA GTAGGGGCAT GGTATAAGGA AATATCATAA TGTAATGAGG 2280 TGGTACTATG ATTAAAAGCC ACTGTTACAT TTCAAAAAAA AAAAAAAAAA ACCATCTAAA 2340 AAAGGTAGGT AGGTAAAAGA ATTATATTAT CAA 2373 (2) INFORMATION FOR SEQ ID NO:2 : (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 651 amino acids (B) TYPE: amino acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: protein (xi) SEQUENCE DESCRIPTION: SEQ ID NO:2 : Gly Gin Asp Phe Gin Asn Phe Cys His Tyr Gin Gly Tyr lie Glu Gly 1 5 10 15 Tyr Pro Lys Ser Val Val Met Val Ser Thr Cys Thr Gly Leu Arg Gly 20 25 30 Val Leu Gin Phe Glu Asn Val Ser Tyr Gly lie Glu Pro Leu Glu Ser 35 40 45 Ser Val Gly Phe Glu His Val lie Tyr Gin Val Lys His Lys Lys Ala 50 55 60 Asp Val Ser Leu Tyr Asn Glu Lys Asp lie Glu Ser Arg Asp Leu Ser 65 70 75 80 Phe Lys Leu Gin Ser Ala Glu Pro Gin Gin Asp Phe Ala Lys Tyr lie 85 90 95 Glu Met His Vαl He Val Glu Lys Gin Leu Tyi Asn His Met Gly Sei 100 105 110 Asp Thr Thr Val Val Ala Gin Lys Val Phe Gin Leu He Gly Leu Thr 115 120 125 Asn Ala He Phe Val Sei Phe Asn He Thi He He Leu Ser Sei Leu 130 135 140 Glu Leu T p He Asp Glu Asn Lys He Ala Thi Thi Gly Glu Ala Asn 145 150 155 160 Glu Leu Leu His Th Phe Leu Arg Tip Lys Thi Sei Tyi Leu Val Leu 165 170 175 Aig PLO HIS Asp Val Ala Phe Leu Leu Vαl Ty Aig Glu Lys ^ei Asn 130 185 190 Tyi Vαl Gly Ala Thi Phe Gin Gly Lys Met Cys Asp Ala Asn Ty Ala 195 200 205 Gly Gly Val Val Leu His Pro Arg Thr He Sei Leu Glu Se Leu Ala 210 215 220 Val He Leu Ala Gin Leu Leu Ser Leu Sei Met Gly He Thi Tyi Asp 225 230 235 " 240 Asp He Asn Lys Cys Gin Cys Ser Gly Ala Val Cys He Met Asn Pro 245 250 255 Glu Ala He His Phe Sei Gly Val Lys He Asn Cys Sei Phe 260 265 270 Glu Asp Phe Ala His Phe He Sei Lys Gin Lys Sei Gin Cys Leu His 275 280 285 Asn Gin P o Aig Leu Asp Pro Phe Phe Lys Gin Gin Ala Val Cys Gly 290 295 300 Asn Ala Lys Leu Glu Ala Gly Glu Glu Cys Asp Cys Gly Thi Glu Gin 305 310 315 320 Asp Cys Ala Leu He Gly Glu Thi Cys Cys Asp He Ala Thi Cys Aig 325 330 335 Phe Lys Ala Gly Ser Asn Cys Ala Glu Gly Pio Cys Cys Glu Asn Cys 340 " 345 350 Leu Phe Met Ser Lys Glu Arg Met Cys Arg Pro ei Phe Glu Glu Cys 355 360 365 Asp Leu Pio Glu Tyr Cys Asn Gly Ser Ser Ala Sei Cys Pro Glu Asn 370 375 330 His Tyr Val Gin Thr Gly His Pro Cys Gly Leu Asn Gin Tip He Cys 385 390 395 400 He Asp Gly Val Cys Met Ser Gly Asp Lys Gin Cys Thr Asp Thr Phe 405 410 415 Gly Lys Glu Val Glu Phe Gly Pro Ser Glu Cys Tyr Ser His Leu Asn 420 425 430 Ser Lys Thr Asp Val Ser Gly Asn Cys Gly He Ser Asp Ser Gly Tyr 435 440 445 Thr Gin Cys Glu Ala Asp Asn Leu Gin Cys Gly Lys Leu He Cys Lys 450 455 460 Tyr Val Gly Lys Phe Leu Leu Gin He Pro Arg Ala Thr He He Tyr 4 * 65 470 475 480 Ala Asn He Ser Gly His Leu Cys He Ala Val Glu Phe Ala Ser Asp 485 490 495 His Ala Asp Ser Gin Lys Met Trp He Lys Asp Gly Thr Ser Cys Gly 500 505 510 Ser Asn Lys Val Cys Arg Asn Gin Arg Cys Val Ser Ser Ser Tyr Leu 515 520 525 Gly Tyr Asp Cys Thr Thr Asp Lys Cys Asn Asp Arg Gly Val Cys Asn 530 535 540 Asn Lys Lys His Cys His Cys Ser Ala Ser Tyr Leu Pro Pro Asp Cys 545 " * 550 555 560 Ser Val Gin Ser Asp Leu Trp Pro Gly Gly Ser He Asp Ser Gly Asn 565 570 575 Phe Pro Pro Val Ala He Pro Ala Arg Leu Pro Glu Arg Arg Tyr He 530 585 590 Glu Asn He Tyr His Ser Lys Pro Met Arg Trp Pro Phe Phe Leu Phe 595 600 605 I. Pro Phe Phe He He Phe Cys Val Leu He Ala He Met Val Lys 610 615 620 Val Asn Phe Gin Arg Lys Lys Trp Arg Thr Glu Asp Tyr Ser Ser Asp 625 630 635 640 Glu Gin Pro Glu Ser Glu Ser Glu Pro Lys Gly 645 650 (2) INFORMATION FOR SEQ ID NO:3 : (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 1768 base pairs (B) TYPE: nucleic acid ■ C) STRANDEDNESS: double (D) TOPOLOGY: linear (HI MOLECULE TYPE: cDNA (xi) SEQUENCE DESCRIPTION: SEQ ID NO:3 : GGCACGAGCG ATTATGTTGG CGCTACCTAT CAAGGGAAGA TGTGTGZVCAA GAACTATGCA 60 GGAGGAGTTG CTTTGCACCC CAAAGCCGTA ACTCTGGAAT CACTTGCAAT TATTTTAGTT 120 CAGCTGCTGA GCCTCAGCAT GGGGCTAGCG TATGACGACG TGAACAAGTG CCAGTGTGGC 130 GTACCTGTCT GCGTGATGAA CCCGGAAGCG CCTCACTCCA GCGGTGTCCG GGCCTTCAGT 240 AACTGCAGCA TGGAGGACTT TTCCAAGTTT ATCACAAGTC AAAGCTCCCA CTGTCTGCAG 300 AACCAGCCAA CGCTACAGCC ATCTTACAAG ATGGCGGTCT GTGGGAATGG AGAGGTGGAA 360 GAAGATGAAA TTTGCGACTG TGGAAAGAAG GGCTGTGCAG AAATGCCCCC GCCATGCTGT 420 AACCCCGACA CCTGTAAGCT GTCAGATGGC TCCGAGTGCT CCAGCGGGAT ATGCTGCAAC 480 TCGTGCAAGC TGAAGCGGAA AGGGGAGGTT TGCAGGCTTG CCCAAGATGA GTGTGATGTC 540 ACAGAGTACT GCAACGGCAC ATCCGAAGTG TGTGAAGACT TCTTTGTTCA AAACGGTCAC 600 CCATGTC CA ATCGCAAGTG GATCTGTATT AACGGCACCT GTCAGAGTGG AGAACAGCAG 660 TGCCAGG^TC TATTTGGCAT CGATGCAGGC TTTGGTTCAA GTGAATGTTT CTGGGAGCTG 720 AATTCCAAGA GCGACATATC TGGGAGCTGT GGAATCTCTG CTGGGGGAT^ CAAGGAATGC 780 CCACCTAATG ZVCCGGATGTG TGGGAAAATA ATATGTAAAT ACCAA^GTGZV AAATATACT 840 AAATTGAGGT CTGCCACTGT TATTTATGCC AATATAAGCG GGCATGTCTG CGTTTCCCTG 900 GAATATCCCC AAGGTCATAA TGAGAGCCAG AAGATGTGGG TGAGAGATGG AACCGTCTGC 960 GGGTCAAATA AGGTTTGCCA GAATCAAAAA TGTGTAGCAG ACACTTTCTT GGGCTATGAT 1020 TGCAACCTGG A AAATGCAA CCACCATGGT GTATGTAATA ACAAGAAGAA CTGCCACTGT 1080 GACCCCA AT ACTTACCTCC AGATTGTAAA AGAATGAAAG ATTCATATCC TGGCGGGAGC 1140 ATTGATAGTG GCAACAAGGA AAGGGCTGAA CCCATCCCTG TACGGCCCTA CATTGCAAGT 1200 CGTTACCGCT CCAAGTCTCC ACGGTGGCCA TTTTTCTTGA TCATCCCTTT CTACGTTGTG 1260 ATCCTTGTCC TGATTGGGAT GCTGGTAAAA GTCTATTCCC AAAGGATGAA ATGGAGAATG 1320 GATGACTTCT CAAGCGAAGA GCAATTTGAA AGTGAAAGTG AATCCAAAGA CTAGTCTGGA 1380 CAGATTCCAC AATGTCACAA GTAATTCTCT TCAGTGGACA GAAAAAAAAG TGGAAAAGAA 1440 AA3CCTVTGC ATTATCTTGC CTGAAAGTCA AGCCTGCATA TCGTGGTCTC CATCAGGCCA 1500 GAAATCATAT CTCTCCATTA CACATGTATG ATACATATGT GTGTATATTA TTCCATAAAT 1560 GATTTACTTG TAAGAAATGA ATGATTATGA ATTTCATATT ATACTTTGAT ATTTTACCCT 1620 ATTTCTGGTA GTCGGTAGTC ATCAATTGTA TTTTCTAGTA GGTACATTAT AGAAAAGGCT 1680 ATAAGAAAAT AAATGTGGTA CCATAATAAT CAATATCATA CAACCACCAT CTAAAAAAGG 1740 TAGGTAGGTA AAAGAATTAT ATTATCAA 1768 (2) INFORMATION FOR SEQ ID NO:4 : (l) SEQUENCE CHARACTERISTICS: (A) LENGTH: 457 ammo acids (B) TYPE, ammo acid (C) STRANDEDNESS: singlp (D) TOPOLOGY- linear (nl MOLECULE TYPE: piotein (xi) SEQUENCE DESCRIPTION: SEQ ID NO:4 : Gly Thr Ser Asp Tyr Val Gly Ala Thr Tyi Gin Gly Lys Met Cys Asp 1 5 10 15 Lys Asn Tyi Ala Gly Gly Val Ala Leu His Pio Lys Ala Vαl Thi Leu 20 25 30 Glu ^ei Leu Al i He He Leu Val Gin Leu Leu Sei Leu S ^ Met Gly 35 40 45 Leu Ala Tyi Asp Asp Val Asn Lys Cys Gin Cys Gly Val Pio Val Cys 50 55 6θ " Val Met Asn P o Glu Ala Pro His Sei Ser Gly Val Aig Ala Phe Sei 65 70 75 80 Asn Cys Sei Met Glu Asp Phe Ser Lys Phe He Thr Sei Gin Sei Sei 85 90 95 His Cys Leu Gin Asn Gin Pro Thi Leu Gin Pio Sei Tyi Lys Met Ala 100 105 110 Val Cys Gly Asn Gly Glu Val Glu Glu Asp Glu He Cys Asp Cys Gly 115 120 125 Lys Lys Gly Cys Ala Glu Met Pro Pro Pio Cys Cys Asn Pio Asp Thi 130 135 140 Cys Lys Leu Sei Asp Gly Ser Glu Cys Sei Sei Gly He Cys Cys Asn 145 150 155 " 160 Sei Cys Lys Leu Lys Aig Lys Gly Glu Val Cys Aig Leu ^la Gin Asp 165 170 175 Glu Cys Asp Val Thr Glu Tyr Cys Asn Gly Thr Ser Glu Val Cys Glu 180 185 190 Asp Phe Phe Val Gin Asn Gly His Pro Cys Asp Asn Arg Lys Trp He 195 200 205 Cvs He Asn Glv Thr Cys Gin Ser Gly Glu Gin Gin Cys Gin Asp Leu 210 " 215 220 Phe Gly He Asp Ala Gly Phe Gly Ser Ser Glu Cys Phe Trp Glu Leu 225 230 235 240 Asn Ser Lys Ser Asp He Ser Gly Ser Cys Gly He Ser Ala Gly Gly 245 250 255 Tyr Lys Glu Cys Pro Pro Asn Asp Arg Met Cys Gly Lys He He Cys 260 265 270 Lys Tyr Gin Ser Glu Asn He Leu Lys Leu Arg Ser Ala Thr Val He 275 280 285 Tyr Ala Asn He Ser Gly His Val Cys Val Ser Leu Glu Tyr Pro Gin 290 295 300 Gly His Asn Glu Ser Gin Lys Met Trp Val Arg Asp Gly Thr Val Cvs 305 310 315 320 Gly Ser Asn Lys Val Cys Gin Asn Gin Lys Cys Val Ala Asp Thr Phe 325 330 335 Leu Gly Tyr Asp Cys Asn Leu Glu Lys Cys Asn His His Gly Val Cys 340 " 345 350 Asn Asn Lvs Lvs Asn Cys His Cys Asp Pro Thr Tyr Leu Pro Pro Asp 355 " 360 ' 365 Cys Lys Arg Met Lys Asp Ser Tyr Pro Gly Gly Ser He Asp Ser Gly 370 375 330 Asn Lys Glu Arg Ala Glu Pro He Pro Val Arg Pro Tyr He Ala Ser 335 390 395 400 Arg Tyr Arg Ser Lys Ser Pro Arg Trp Pro Phe Phe Leu He He Pro 405 410 415 Phe Tyr Val Val He Leu Val Leu He Gly Met Leu Val Lys Val Tyr 420 425 430 Ser Gin Arg Met Lys Trp Arg Met Asp Asp Phe Ser Ser Glu Glu Gin 435 440 445 Phe Glu Ser Glu Ser Glu Ser Lys Asp 450 455 (2) INFORMATION FOR SEQ ID NO:5: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 2553 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS : double (D) TOPOLOGY: linear (ii < MOLECULE TYPE: cDNA (ix ■ FEATURE: (A) NAME/KEY: CDS (B) LOCATION: 17..2221 (xi' SEQUENCE DESCRIPTION: SEQ ID NO: 5 : TGAGGAGGAC CAGCGC ATG CGG CTC ATC TTG CTT CTA CTG AGT GGG CTG 49 Met Arg Leu He Leu Leu Leu Leu Ser Gly Leu 1 5 10 AGT GAA CTT GGC GGC CTT AGC CAG TCC CAA ACA GAA GGC ACT CGT GAG 97 Ser Glu Leu Gly Gly Leu Ser Gin Ser Gin Thr Glu Gly Thr Arg Glu 15 20 25 AAA TTA CAC GTG CAA GTC ACA GTG CCA GAG AAA ATC CGG TCC GTC ACA 145 Lys Leu His Val Gin Val Thr Val Pro Glu Lys He Arg Ser Val Thr 30 35 40 AGC AAT GGC TAC GAA ACA CAG GTG ACC TAC AAT CTC AAA ATC GAA GGG 193 Ser Asn Gly Tyr Glu Thr Gin Val Thr Tyr Asn Leu Lys He Glu Gly 45 50 55 AAA AC.-. TAC ACC TTG GAC CTA ATG CAA AAA CCG TTC TTG CCT CCC AAC 241 Lys Thr Tyr Thr Leu Asp Leu Met Gin Lys Pro Phe Leu Pro Pro Asn 60 65 70 75 TTT AGA GTA TAC AGT TAT GAC AAC GCA GGA ATC ATG AGG TCT CTT GAG 239 Phe Arg Val Tyr Ser Tyr Asp Asn Ala Gly He Met Arg Ser Leu Glu 30 85 90 CAG AAG TTT CAG AAT ATC TGC TAC TTC CAA GGA TAC ATT GAA GGT TAT 337 Gin Lys Phe Gin Asn He Cys Tyr Phe Gin Gly Tyr He Glu Gly Tyr 95 100 105 CCA AAT TCT ATG GTG ATT GTT AGC ACA TGT ACT GGA CTG AGG GGT TTT 385 Pro Asr. Ser Met Val He Val Ser Thr Cys Thr Gly Leu Arg Gly Phe 110 115 120 CTC CAA TTT GGA AAC GTT AGC TAT GGA ATT GAA CCT CTG GAA TCT TCC 433 Leu Gin Phe Gly Asn Val Ser Tyr Gly He Glu Pro Leu Glu Ser Ser 125 130 135 AGT GGT TTT GAA CAC GTG ATC TAC CAA GTG GAA CCT GAG AAA GGA GGT 481 Ser Gly Phe Glu His Val He Tyr Gin Val Glu Pro Glu Lys Gly Gly 140 145 150 155 GCA TTA CTC TAC GCC GAG AAG GAT ATC GAT TTA AGA GAC TCG CAG TAT 529 Ala Leu Leu Tyr Ala Glu Lys Asp He Asp Leu Arg Asp Ser Gin Tyr 160 165 170 AAG ATA CGA AGT ATC AAG CCA CAG CGG ATC GTC TCT CAC TAT TTG GAA 577 Lys He Arg Ser He Lys Pro Gin Arg He Val Ser His Tyr Leu Glu 175 180 185 ATA CAT ATT GTC GTT GAA AAG CAA ATG TTT GAG CAT ATC GGG GCT GAT 625 He His He Val Val Glu Lys Gin Met Phe Glu His He Gly Ala Asp 190 195 200 ACA GCC ATT GTC ACT CAA AAG ATT TTC CAG TTG ATT GGA CTG GCA AAT 673 Thr Ala He Val Thr Gin Lys He Phe Gin Leu He Gly Leu Ala Asn 205 210 215 GCT ATC TTT GCC CCC TTT AAT CTT ACA GTA ATT CTG TCT TCC CTG GAA 721 Ala He Phe Ala Pro Phe Asn Leu Thr Val He Leu Ser Ser Leu Glu 220 225 230 235 TTT TGG ATG GAT GAA AAC AAA ATC TTG ACC ACA GGC GAT GCT AAC AAG 769 Phe Trp Met Asp Glu Asn Lys He Leu Thr Thr Gly Asp Ala Asn Lys 240 245 250 TTG CTC TAC AGG TTC CTG AAG TGG AAA CAG TCG TAC CTT GTT CTG CGA 817 Leu Leu Tyr Arg Phe Leu Lys Trp Lys Gin Ser Tyr Leu Val Leu Arg 255 260 265 CCA CAT GAT ATG GCG TTT TTA CTC GTC TAC AGG AAC ACT ACC GAT TAT 865 Pro Hi- Asp Met Ala Phe Leu Leu Val Tyr Arg Asn Thr Thr Asp Tyr 270 275 280 GTT GGC GCT ACC TAT CAA GGG AAG ATG TGT GAC AAG AAC TAT GCA GGA 913 Val Gly Ala Thr Tyr Gin Gly Lys Met Cys Asp Lys Asn Tyr Ala Gly 28: 290 295 GGA GTT GCT TTG CAC CCC AAA GCC GTA ACT CTG GAA TC'A CTT GCA ATT 961 Gly Val Ala Leu His Pro Lys Ala Val Thr Leu Glu Ser Leu Ala He 300 305 310 315 ATT TTA GTT CAG CTG CTG AGC CTC AGC ATG GGG CTA GCG TAT GAC GAC 1009 He Lev. Val Gin Leu Leu Ser Leu Ser Met Gly Leu Ala Tyr Asp Asp 320 325 330 GTG AAC AAG TGC CAG TGT GGC GTA CCT GTC TGC GTG ATG AAC CCG GAA 1057 Val Asr. Lys Cys Gin Cys Gly Val Pro Val Cys Val Met Asn Pro Glu 335 340 345 GCG CCT CAC TCC AGC GGT GTC CGG GCC TTC AGT AAC TGC AGC ATG GAG 1105 Ala Pre His Ser Ser Gly Val Arg Ala Phe Ser Asn Cys Ser Met Glu 350 355 360 GAC TTT TCC AAG TTT ATC ACA AGT CAA AGC TCC CAC TGT CTG CAG AAC 1153 Asp Phe Sei Lys Phe He Thi Sei Gin Sei Sei His Cys Leu Gin Asn 365 370 375 CAG CCA ACG CTA CAG CCA TCT TAC AAG ATG GCG GTC TGT GGG AAT GGA 1201 Gin Pro Thr Leu Gin Pro Ser Tyr Lys Met Ala Val Cys Gly Asn Gly 380 385 390 395 GAG GTG GAA GAA GAT GAA ATT TGC GAC TGT GGA AAG AAG GGC TGT GCA 1249 Glu Val Glu Glu Asp Glu He Cys Asp Cys Gly Lys Lys Gly Cys Ala 400 405 410 GAA ATG CCC CCG CCA TGC TGT AAC CCC GAC ACC TGT AAG CTG TCA GAT 1297 Glu Met Pio Pio Pio Cys Cys Asn Pio Asp Thi Cys Lys Leu Sei Asp 415 420 425 GGC TCC GAG TGC TCC AGC GGG ATA TGC TGC AAC TCG TGC AAG CTG AAG 1345 Gly Sei Glu Cys Sei Sei Gly He Cys Cys Asn Sei Cys Lys Leu Lys 430 435 440 CGG AAA GGG GAG GTT TGC AGG CTT GCC CAA GAT GAG TGT GAT GTC ACA 1393 Aig Lys Gly Glu Val Cys Aig Leu Ala Gin Asp Glu Cys Asp Val Thi 445 450 455 GAG TAC TGC AAC GGC ACA TCC GAA GTG TGT GAA GAC TTC TTT GTT CAA 1441 Glu Tyi Cys Asn Gly Thi Sei Glu Val Cys Glu Asp Phe Phe Val Gin 460 " " 465 470 475 AAC GGT CAC CCA TGT GAC AAT CGC AAG TGG ATC TGT ATT AAC GGC ACC 1489 Asn Gly His Pio Cys Asp Asn Aig Lys Tip He Cys He Asn Gly Thi 480 485 490 TGT CAG AGT GGA GAA CAG CAG TGC CAG GAT CTA TTT GGC ATC GAT GCA 1537 Cys Gin Sei Gly Glu Gin Gin Cys Gin Asp Leu Phe Gly He As}. Ala 495 500 505 GGC TTT GGT TCA AGT GAA TGT TTC TGG GAG CTG AAT TCC AAG AGC GAC 1535 Gly Phe Gly Sei Sei Glu Cys Phe Trp Glu Leu Asn Sei Lys Sei Asp 510 515 520 ATA TCT GGG AGC TGT GGA ATC TCT GCT GGG GGA TAC AAG GAA TGC CCA 1633 He Sei Gly Ser Cys Gly He Ser Ala Gly Gly Tyi Lys Glu Cys Pio 525 530 535 CCT AAT GAC CGG ATG TGT GGG AAA ATA ATA TGT AAA TAC CAA AGT GAA 1681 Pio Asn Asp Aig Met Cys Gly Lys He He Cys Lys Tyi Gin Se Glu 540 545 550 555 AAT ATA CTA AAA TTG AGG TCT GCC ACT GTT ATT TAT GCC AAT ATA AGC 1729 Asn He Leu Lys Leu Aig Sei Ala Thr Val He Tyi Ala Asn He Sei 560 565 570 GGG CAT GTC TGC GTT TCC CTG GAA TAT CCC CAA GGT CAT AAT GAG AGC 1777 Gly H s Val Cys Val Sei Leu Glu Tyr Pro Gin Gly His Asn Glu Ser 575 580 535 CAG AAG ATG TGG GTG AGA GAT GGA ACC GTC TGC GGG TCA AAT AAG GTT 1825 Gin Lys Met Trp Val Arg Asp Gly Thr Val Cys Gly Ser Asn Lys Val 590 595 600 TGC CAG AAT CAA AAA TGT GTA GCA GAC ACT TTC TTG GGC TAT GAT TGC 1873 Cys Gin Asn Gin Lys Cys Val Ala Asp Thr Phe Leu Gly Tyr Asp Cys 605 610 615 AAC CTG GAA AAA TGC AAC CAC CAT GGT GTA TGT AAT AAC AAG AAG AAC 1921 Asn Leu Glu Lys Cys Asn His His Gly Val Cys Asn Asn Lys Lys Asn 620 " 625 630 635 TGC CAC TGT GAC CCC ACA TAC TTA CCT CCA GAT TGT AAA AGA ATG AAA 1969 Cys His Cys Asp Pro Thr Tyr Leu Pro Pro Asp Cys Lys Arg Met Lys 640 645 650 GAT TCA TAT CCT GGC GGG AGC ATT GAT AGT GGC AAC AAG GAA AGG GCT 2017 Asp Ser Tvr Pro Gly Gly Ser He Asp Ser Gly Asn Lys Glu Arg Ala 655 660 665 GAA CCC ATC CCT GTA CGG CCC TAC ATT GCA AGT CGT TAC CGC TCC AAG 2065 Glu Pro He Pro Val Arg Pro Tyr He Ala Ser Arg Tyr Arg Ser Lvs 670 675 680 TCT CCA CGG TGG CCA TTT TTC TTG ATC ATC CCT TTC TAC GTT GTG ATC 2113 Ser Pro Arg Trp Pro Phe Phe Leu He He Pro Phe Tyr Val Val He 685 690 695 CTT GTC CTG ATT GGG ATG CTG GTA AAA GTC TAT TCC CAA AGG ATG AAA 2161 Leu Val Leu He Gly Met Leu Val Lys Val Tyr Ser Gin Arg Met Lys 700 705 710 715 TGG AGA ATG GAT GAC TTC TCA AGC GAA GAG CAA TTT GAA AGT GAA AGT 2209 Trp Arg Met Asp Asp Phe Ser Ser Glu Glu Gin Phe Glu Ser Glu Ser 720 725 730 GAA TCC AAA GAC TAGTCTGGAC AGATTCCACA ATGTCACAAG TAATTCTCTT 2261 Glu Ser Lys Asp 735 C.AGTGGAC.AG AAAAAAAAGT GGAAAAGAAA AGCCT.ATGCA TT.ATCTTGCC TGAAAGTCAA 2321 GCCTGC.AT.AT CGTGGTCTCC ATCAGGCCAG AAATCATATC TCTCC.ATTAC ACATGT.ATGA 2381 TAC.ATATGTG TGTATATT.AT TCCATAAATG ATTTACTTGT AAGAAATG.AA TGATT.ATGAA 2441 TTTCATATTA TACTTTGATA TTTTACCCTA TTTCTGGTAG TCGGTAGTCA TCAATTGTAT 2501 TTTCTAGTAG GTACATTATA GAAAAGGCTA TAAGAAAATA AATGTGGTAC CA 2553 (2) INFORMATION FOR SEQ ID NO: 6 : (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 735 amino acids (B) TYPE: amino acid (D) TOPOLOGY: linear (11) MOLECULE TYPE: protein (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 6 : Met Arg Leu He Leu Leu Leu Leu Ser Gly Leu Ser Glu Leu Gly Gly 1 5 10 15 Leu Ser Gin Ser Gin Thr Glu Gly Thr Arg Glu Lys Leu His Val Gin 20 25 30 Val Thr Val Pro Glu Lys He Arg Ser Val Thr Ser Asn Gly Tyr Glu 35 40 45 Thi Gin Val Thr Tyr Asn Leu Lys He Glu Gly Lys Thr Tyr Thi Leu 50 55 60 Asp Leu Met Gin Lys Pro Phe Leu Pro Pro Asn Phe Arg Val Tyr Ser 65 70 75 80 Tyr Asp Asn Ala Gly He Met Arg Ser Leu Glu Gin Lys Phe Gin Asn 35 90 95 He Cys Tyr Phe Gin Gly Tyr He Glu Gly Tyr Pro Asn Ser Met Val 100 105 110 He Val Sei Thr Cys Thr Gly Leu Arg Gly Phe Leu Gin Phe Gly Asn 115 120 125 Val Sei Tyr Gly He Glu Pro Leu Glu Ser Ser Ser Gly Phe Glu His 130 135 140 Val He Tyi Gin Val Glu Pro Glu Lys Gly Gly Ala Leu Leu Tyr Ala 145 150 155 160 Glu Lys Asp He Asp Leu Arg Asp Ser Gin Tyr Lys He Arg Ser He 165 170 175 Lys Pro Gin Arg He Val Ser His Tyr Leu Glu He His He Val Val 180 185 190 Glu Lys Gin Met Phe Glu His He Gly Ala Asp Thr Ala He Val Thr 195 200 205 Gin Lys He Phe Gin Leu He Gly Leu Ala Asn Ala He Phe Ala Pro 210 215 220 Phe Asn Leu Thr Val He Leu Ser Ser Leu Glu Phe Trp Met Asp Glu 225 230 235 240 Asn Lys lie Leu Thr Thr Gly Asp Ala Asn Lys Leu Leu Tyr Arg Phe 245 250 255 Leu Lys Trp Lys Gin Ser Tyr Leu Val Leu Arg Pro His Asp Met Ala 260 265 270 Phe Leu Leu Val Tyr Arg Asn Thr Thr Asp Tyr Val Gly Ala Thr Tyr 275 280 285 Gin Gly Lys Met Cys Asp Lys Asn Tyr Ala Gly Gly Val Ala Leu His 290 " 295 300 Pro Lys Ala Val Thr Leu Glu Ser Leu Ala He He Leu Val Gin Leu 305 310 315 320 Leu Ser Leu Ser Met Gly Leu Ala Tyr Asp Asp Val Asn Lys Cys Gin 325 330 335 Cys Gly Val Pro Val Cys Val Met Asn Pro Glu Ala Pro His Ser Ser 340 345 350 Gly Val Arg Ala Phe Ser Asn Cys Ser Met Glu Asp Phe Ser Lys Phe 355 360 365 He Thr Ser Gin Ser Ser His Cys Leu Gin Asn Gin Pro Thr Leu Gin 370 375 380 Pro Ser Tyr Lys Met Ala Val Cys Gly Asn Gly Glu Val Glu Glu Asp 385 390 395 400 Glu He Cys Asp Cys Gly Lys Lys Gly Cys Ala Glu Met Pro Pro Pro 405 410 415 Cys Cys Asn Pro Asp Thr Cys Lys Leu Ser Asp Gly Ser Glu Cys Ser 420 425 430 Ser Gly He Cys Cys Asn Ser Cys Lys Leu Lys Arg Lys Gly Glu Val 435 440 445 Cys Arg Leu Ala Gin Asp Glu Cys Asp Val Thr Glu Tyr Cys Asn Gly 450 455 460 Thr Ser Glu Val Cys Glu Asp Phe Phe Val Gin Asn Gly His Pro Cys 465 470 475 480 Asp Asn Arg Lys Trp He Cys He Asn Gly Thr Cys Gin Ser Gly Glu 485 490 495 Gin Gin Cys Gin Asp Leu Phe Gly He Asp Ala Gly Phe Gly Ser Ser 500 505 510 Glu Cys Phe Trp Glu Leu Asn Ser Lys Ser Asp He Ser Gly Ser Cys 515 520 525 Gly He Ser Ala Gly Gly Tyr Lys Glu Cys Pro Pro Asn Asp Arg Met 530 535 540 Cys Gly Lys He He Cys Lys Tyr Gin Ser Glu Asn He Leu Lys Leu 545 " " 550 " 555 560 Arg Ser Ala Thr Val He Tyr Ala Asn He Ser Gly His Val Cys Val 565 570 575 Ser Leu Glu Tyr Pro Gin Gly His Asn Glu Ser Gin Lys Met Trp Val 580 585 590 Arg Asp Gly Thr Val Cys Gly Ser Asn Lys Val Cys Gin Asn Gin Lys 595 600 605 Cys Val Ala Asp Thr Phe Leu Gly Tyr Asp Cys Asn Leu Glu Lys Cys 610 615 620 Asn His His Gly Val Cys Asn Asn Lys Lys Asn Cys His Cys Asp Pro 625 630 635 640 Thr Tyr Leu Pro Pro Asp Cys Lys Arg Met Lys Asp Ser Tyr Pro Gly 645 650 655 Gly Ser He Asp Ser Gly Asn Lys Glu Arg Ala Glu Pro He Pro Val 660 665 670 Arg Pro Tyr He Ala Ser Arg Tyr Arg Ser Lys Ser Pro Arg Trp Pro 675 680 685 Phe Phe Leu He He Pro Phe Tyr Val Val He Leu Val Leu He Gly 690 695 700 Met Leu Val Lys Val Tyr Ser Gin Arg Met Lys Trp Arg Met Asp Asp 705 710 715 720 Phe Ser Ser Glu Glu Gin Phe Glu Ser Glu Ser Glu Ser Lys Asp 725 730 735 (2) INFORMATION FOR SEQ ID NO:7 : (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 2650 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: double (D) TOPOLOGY: linear (ii) MOLECULE TYPE: cDNA (ixi 'FEATURE: (A) NAME/KEY: CDS (B) LOCATION: 72..2273 (xi) SEQUENCE DESCRIPTION: SEQ ID .NO:7 : CATCTCGCAC TTCCAACTGC CCTGTAACCA CCAACTGCCC TTATTCCGGC TGGGACCCAG 60 GACTTCAAGC C ATG TGG GTC TTG TTT CTG CTC AGC GGG CTC GGC GGG CTG 110 Met Trp Val Leu Phe Leu Leu Ser Gly Leu Gly Gly Leu 740 745 CGG ATG GAC AGT AAT TTT GAT AGT TTA CCT GTG CAA ATT ACA GTT CCG 158 Aig Met Asp Ser Asn Phe Asp Ser Leu Pro Val Gin He Thi Vαl Pio 750 755 760 GAG AAA ATA CGG TCA ATA ATA AAG GAA GGA ATT GAA TCG CAG GCA TCC 206 Glu Lys He A g Sei He He Lys Glu Gly He Glu Sei Gin Ala Sei 765 770 775 780 TAC AAA ATT GTA ATT GAA GGG AAA CCA TAT ACT GTG AAT TTA ATG CAA 254 Tyi Lys He Vαl He Glu Gly Lys Pro Tyr Thi Val Asn Leu Met Gin 785 790 795 AAA AAC TTT TTA CCC CAT AAT TTT AGA GTT TAC AGT TAT AGT GGC ACA 302 Lys Asn Phe Leu Pio His Asn Phe Aig Val Tyi Sei Tyi Sei Gly Thi 800 305 810 GGA ATT ATG AAA CCA CTT GAC CAA GAT TTT CAG AAT TTC TGC CAC TAC 350 Gly He Met Lys Pro Leu Asp Gin Asp Phe Gin Asn Phe Cys His Tyi 815 820 325 CAA GGG TAT ATT GAA GGT TAT CCA AAA TCT GTG GTG ATG GTT AGC ACA 398 Gin Gly Tyi He Glu Gly Tyr Pro Lys Ser Val Val Met Vαl Ser Th 830 835 840 TGT ACT GGA CTC AGG GGC GTA CTA CAG TTT GAA AAT GTT AGT TAT GGA 446 Cys Thi Gly Leu Aig Gly Val Leu Gin Phe Glu Asn Val Sei Tyi Gly 845 850 855 860 ATA GAA CCC CTG GAG TCT TCA GTT GGC TTT GAA CAT GTA ATT TAC CAA 494 He Glu Pio Leu Glu Sei Sei Val Gly Phe Glu His VH He Tyi Gin 865 370 375 GTA AAA CAT AAG AAA GCA GAT GTT TCC TTA TAT AAT GAG AAG GAT ATT 542 V__l Lys His Lys Lys Ala Asp Val Sei Leu Tyi Asn Glu Lys Asp He 880 " 385 890 GAA TCA AGA GAT CTG TCC TTT AAA TTA CAA AGC GCA GAG CCA CAG CAA 590 Glu Sei A g Asp Leu Sei Phe Lys Leu Gin Sei Ala Glu Pio Gin Gin 395 900 905 GAT TTT GCA AAG TAT ATA GAA ATG CAT GTT ATA GTT GAA AAA CAA TTG 633 Asp Phe Ala Lvs Tyi He Glu Met His Val He Val Glu Lys Gin Leu 910 915 920 TAT AAT CAT ATG GGG TCT GAT ACA ACT GTT GTC GCT CAA AAA GTT TTC 686 Ty Asn His Met Gly S- Asp Thi Thi Val Val Ala Gin Lys Val Phe 925 930 935 940 CAG TTG ATT GGA TTG ACG AAT GCT ATT TTT GTT TCA TTT AAT ATT ACA 734 Gin Leu He Gly Leu Thi Asn Ala He Phe Vαl Sei phe Asn He Thi 945 950 955 ATT ATT CTG TCT TCA TTG GAG CTT TGG ATA GAT GAA AAT AAA ATT GCA 732 He He Leu Ser Sei Leu Glu Leu Trp He Asp Glu Asn Lys He Ala 960 965 970 ACC ACT GGA GAA GCT AAT GAG TTA TTA CAC ACA TTT TTA AGA TGG AAA 330 Thr Thr Gly Glu Ala Asn Glu Leu Leu His Thr Phe Leu Arg Trp Lys 975 980 985 ACA TCT TAT CTT GTT TTA CGT CCT CAT GAT GTG GCA TTT TTA CTT GTT 878 Thr Ser Tyr Leu Val Leu Arg Pro His Asp Val Ala Phe Leu Leu Val 990 995 1000 TAC AGA GAA AAG TCA AAT TAT GTT GGT GCA ACC TTT CAA GGG AAG ATG 926 Tyr Arg Glu Lys Ser Asn Tyr Val Gly Ala Thr Phe Gin Gly Lys Met 1005 " 1010 1015 1020 TGT GAT GCA AAC TAT GCA GGA GGT GTT GTT CTG CAC CCC AGA ACC ATA 974 Cys Asp Ala Asn Tyr Ala Gly Gly Val Val Leu His Pro Arg Thr He 1025 1030 1035 AGT CTG GAA TCA CTT GCA GTT ATT TTA GCT CAA TTA TTG AGC CTT AGT 1022 Ser Leu Glu Ser Leu Ala Val He Leu Ala Gin Leu Leu Ser Leu Ser 1040 ' 1045 1050 ATG GGG ATC ACT TAT GAT GAC ATT AAC AAA TGC CAG TGC TCA GGA GCT 1070 Met Gly He Thr Tyr Asp Asp He Asn Lys Cys Gin Cys Ser Gly Ala 1055 1060 1065 GTC TGC ATT ATG AAT CCA GAA GCA ATT CAT TTC AGT GGT GTG AAG ATC 1118 Val Cys He Met Asn Pro Glu Ala He His Phe Sei Gly Val Lys He 1070 1075 1030 TTT AGT AAC TGC AGC TTC GAA GAC TTT GCA CAT TTT ATT TCA AAG CAG 1166 Phe Sei Asn Cys Ser Phe Glu Asp Phe Ala His Phe He Ser Lys Gin 1085 1090 1095 1100 AAG TCC CAG TGT CTT CAC AAT CAG CCT CGC TTA GAT CCT TTT TTC AAA 1214 Lys Sei Gin Cys Leu His Asn Gin Pro Arg Leu Asp Pro Phe Phe Lys 1105 1110 1115 CAG CAA GCA GTG TGT GGT AAT GCA AAG CTG GAA GCA GGA GAG GAG TGT 1262 Gin Gin Ala Val Cys Gly Asn Ala Lys Leu Glu Ala Gly Glu Glu Cys 1120 1125 1130 GAC TGT GGG ACT GAA CAG GAT TGT GCC CTT ATT GGA GAA ACA TGC TGT 1310 Asp Cys Gly Thr Glu Gin Asp Cys Ala Leu He Gly Glu Thr Cys Cys 1135 1140 1145 GAT ATT GCC ACA TGT AGA TTT AAA GCC GGT TCA AAC TGT GCT GAA GGA 1353 Asp He Ala Thr Cys Arg Phe Lys Ala Gly Ser Asn Cys Ala Glu Gly 115: 1155 1160 CCA TGC TGC GAA AAC TGT CTA TTT ATG TCA AAA GAA AGA ATG TGT AGG 1406 Pro Cys Cys Glu Asn Cys Leu Phe Met Ser Lys Glu Arg Met Cys Arg 1165 " 1170 1175 1180 CCT TCC TTT GAA GAA TGC GAC CTC CCT GAA TAT TGC AAT GGA TCA TCT 1454 Pro Ser Phe Glu Glu Cys Asp Leu Pro Glu Tyr Cys Asn Gly Ser Ser 1185 1190 1195 GCA TCA TGC CCA GAA AAC CAC TAT GTT CAG ACT GGG CAT CCG TGT GGA 1502 Ala Ser Cys Pro Glu Asn His Tyr Val Gin Thr Gly His Pro Cys Gly 1200 1205 1210 CTG AAT CAA TGG ATC TGT ATA GAT GGA GTT TGT ATG AGT GGG GAT AAA 1550 Leu Asn Gin Trp He Cys He Asp Gly Val Cys Met Ser Gly Asp Lys 1215 1220 1225 CAA TGT ACA GAC ACA TTT GGC AAA GAA GTA GAG TTT GGC CCT TCA GAA 1598 Gin Cys Thr Asp Thr Phe Gly Lys Glu Val Glu Phe Gly Pro Ser Glu 1230 1235 1240 TGT TAT TCT CAC CTT AAT TCA AAG ACT GAT GTA TCT GGA AAC TGT GGT 1646 Cys Tyr Ser His Leu Asn Ser Lys Thr Asp Val Ser Gly Asn Cys Gly 1245 1250 1255 1260 ATA AGT GAT TCA GGA TAC ACA CAG TGT GAA GCT G.AC AAT CTG CAG TGC 1694 He Ser Asp Ser Gly Tyr Thr Gin Cys Glu Ala Asp Asn Leu Gin Cys 1265 1270 1275 GGA AAA TTA ATA TGT AAA TAT GTA GGT AAA TTT TTA TTA CAA ATT CCA 1742 Gly Lys Leu He Cys Lys Tyr Val Gly Lys Phe Leu Leu Gin He Pro 1280 1285 1290 AGA GCC ACT ATT ATT TAT GCC AAC ATA AGT GGA CAT CTC TGC ATT GCT 1790 Arg Ala Thr He He Tyr Ala Asn He Ser Gly His Leu Cys He Ala 1295 1300 1305 GTG GAA TTT GCC AGT GAT CAT GCA GAC AGC CAA AAG ATG TGG ATA AAA 1838 Val Glu Phe Ala Ser Asp His Ala Asp Ser Gin Lys Met Trp He Lys 13lv 1315 1320 GAT GGA ACT TCT TGT GGT TCA AAT AAG GTT TGC AGG AAT CAA AGA TGT 1886 Asp Gly Thr Ser Cys Gly Ser Asn Lys Val Cys Arg Asn Gin Arg Cys 1325 " 1330 1335 1340 GTG AGT TCT TCA TAC TTG GGT TAT GAT TGT ACT ACT GAC AAA TGC AAT 1934 Val Ser Ser Ser Tyr Leu Gly Tyr Asp Cys Thr Thr Asp Lys Cys Asn 1345 1350 1355 GAT AGA GGT GTA TGC AAT AAC AAA AAG CAC TGT CAC TGT AGT GCT TCA 1982 Asp Arg Gly Val Cys Asn Asn Lys Lys His Cys His Cys Ser Ala Ser 1360 1365 1370 TAT TTA CCT CCA GAT TGC TCA GTT CAA TCA GAT CTA TGG CCT GGT GGG 2030 Tyr Leu Pro Pro Asp Cys Ser Val Gin Ser Asp Leu Trp Pro Gly Gly 1375 1380 1335 AGT ATT G.AC AGT GGC AAT TTT CCA CCT GTA GCT ATA CCA GCC AGA CTC 2078 Ser He Asp Ser Gly Asn Phe Pro Pro Val Ala He Pro Ala Arg Leu 1390 1395 1400 CCT GAA AGG CGC TAC ATT GAG AAC ATT TAC CAT TCC AAA CCA ATG AGA ' 2126 Pro Glu Arg Arg Tyr He Glu Asn He Tyr His Ser Lys Pro Met Arg 1405 1410 1415 1420 TGG CCA TTT TTC TTA TTC ATT CCT TTC TTT ATT ATT TTC TGT GTA CTG 2174 Trp Pro Phe Phe Leu Phe He Pro Phe Phe He He Phe Cys Val Leu 1425 1430 1435 ATT GCT ATA ATG GTG AAA GTT AAT TTC CAA AGG AAA AAA TGG AGA ACT 2222 He Ala He Met Val Lys Val Asn Phe Gin Arg Lys Lys Trp Arg Thr 1440 1445 1450 GAG GAC TAT TCA AGC GAT GAG CAA CCT GAA AGT GAG AGT GAA CCT AAA 2270 Glu Asp Tyr Ser Ser Asp Glu Gin Pro Glu Ser Glu Ser Glu Pro Lys 1455 1460 1465 GGG TAGTCTGGAC AACAGAGATG CCATGATATC ACTTCTTCTA GAGTAATTAT 2323 Gly CTGTGATGGA TGGACACAAA AAAATGGAAA GAAAAGAATG TACATTACCT GGTTTCCTGG 2383 GATTCAAACC TGCATATTGT GATTTTAATT TGACCAGAAA ATATGATATA TATGTATAAT 2443 TTCACAGATA ATTTACTTAT TTAAAAATGC ATGATAATGA GTTTTACATT ACAAATTTCT 2503 GTTTTTTTAA AGTTATCTTA CGCTATTTCT GTTGGTTAGT AGACACTAAT TCTGTCAGTA 2563 GGGGCATGGT ATAAGGAAAT ATCATAATGT AATGAGGTGG TACTATGATT AAAAGCCACT 2623 GTTACATTTC AAAAAAAAAA AAAAAAA 2650 (2) INFORMATION FOR SEQ ID NO:8: (l) SEQUENCE CHARACTERISTICS: (A) LENGTH: 734 amino acids (B) TYPE: ammo acid (D) TOPOLOGY: linear (ιι) MOLECULE TYPE: protein (XI) SEQUENCE DESCRIPTION: SEQ ID NO:8: Met Trp Val Leu Phe Leu Leu Ser Gly Leu Gly Gly Leu Arg Met Asp 1 5 10 15 Ser Asn Phe Asp Ser Leu Pro Val Gin He Thr Val Pro Glu Lys He 20 25 30 Arg Ser He He Lys Glu Gly He Glu Ser Gin Ala Ser Tyr Lys He 35 40 45 Val He Glu Gly Lys Pro Tyr Thr Val Asn Leu Met Gin Lys Asn Phe 50 55 60 Leu Pro His Asn Phe Arg Val Tyr Ser Tyr Ser Gly Thr Gly He Met 65 70 75 80 Lys Pro Leu Asp Gin Asp Phe Gin Asn Phe Cys His Tyr Gin Gly Tyr 85 90 95 He Glu Gly Tyr Pro Lys Ser Val Val Met Val Ser Thr Cys Thr Gly 100 105 110 Leu Arg Gly Val Leu Gin Phe Glu Asn Val Ser Tyr Gly He Glu Pro 115 120 125 Leu Glu Ser Ser Val Gly Phe Glu His Val He Tyr Gin Val Lys His 130 135 140 Lys Lys Ala Asp Val Ser Leu Tyr Asn Glu Lys Asp He Glu Ser Arg 145 150 155 160 Asp Leu Ser Phe Lys Leu Gin Ser Ala Glu Pro Gin Gin Asp Phe Ala 165 170 175 Lys Tyr He Glu Met His Val He Val Glu Lys Gin Leu Tyr Asn His 130 185 190 Met Gly Ser Asp Thr Thr Val Val Ala Gin Lys Val Phe Gin Leu He 195 200 205 Gly Leu Thr Asn Ala He Phe Val Ser Phe Asn He Thr He He Leu 210 215 220 Ser Ser Leu Glu Leu Trp He Asp Glu Asn Lys He Ala Thr Thr Gly 225 230 235 240 Glu Ala Asn Glu Leu Leu His Thr Phe Leu Arg Trp Lys Thr Ser Tyr 245 250 255 Leu Val Leu Arg Pro His Asp Val Ala Phe Leu Leu Val Tyr Arg Glu 260 265 270 Lys Ser Asn Tyr Val Gly Ala Thr Phe Gin Gly Lys Met Cys Asp Ala 275 280 235 Asn Tyr Ala Gly Gly Val Val Leu His Pro Arg Thr He Ser Leu Glu 290 295 300 Ser Leu Ala Val He Leu Ala Gin Leu Leu Ser Leu Ser Met Gly He 305 310 315 320 Thr Tyr Asp Asp He Asn Lys Cys Gin Cys Ser Gly Ala Val Cys He 325 330 335 Met Asn Pro Glu Ala He His Phe Ser Gly Val Lys He Phe Ser Asn 340 345 350 Cys Ser Phe Glu Asp Phe Ala His Phe He Ser Lys Gin Lys Ser Gin 355 360 365 Cys Leu His Asn Gin Pro Arg Leu Asp Pro Phe Phe Lys Gin Gin Ala 370 375 380 Val Cys Gly Asn Ala Lys Leu Glu Ala Gly Glu Glu Cys Asp Cys Gly 385 390 395 400 Thr Glu Gin Asp Cvs Ala Leu He Gly Glu Thr Cys Cys Asp He Ala 405 410 415 Thr Cys Arg Phe Lys Ala Gly Ser Asn Cys Ala Glu Gly Pro Cys Cys 420 425 430 Glu Asn Cys Leu Phe Met Ser Lys Glu Arg Met Cys Arg Pro Ser Phe 435 440 445 Glu Glu Cys Asp Leu Pro Glu Tyr Cys Asn Gly Ser Ser Ala Ser Cys 450 455 460 Pro Glu Asn His Tyr Val Gin Thr Gly His Pro Cys Gly Leu Asn Gin 465 470 475 430 Trp He Cys He Asp Gly Val Cys Met Ser Gly Asp Lys Gin Cys Thr 485 490 495 Asp Thr Phe Gly Lys Glu Val Glu Phe Gly Pro Ser Glu Cys Tyr Ser 500 505 510 His Leu Asn Ser Lys Thr Asp Val Ser Gly Asn Cys Gly He Ser Asp 515 520 525 Ser Gly Tyr Thr Gin Cys Glu Ala Asp Asn Leu Gin Cys Gly Lys Leu 530 535 540 He Cys Lys Tyr Val Gly Lys Phe Leu Leu Gin He Pro Arg Ala Thr 545 " 550 555 560 He He Tyr Ala Asn He Ser Gly His Leu Cys He Ala Val Glu Phe 565 570 575 Ala Ser Asp His Ala Asp Ser Gin Lys Met Trp He Lys Asp Gly Thr 580 585 590 Ser Cys Gly Ser Asn Lys Val Cys Arg Asn Gin Arg Cys Val Ser Ser 595 600 605 Ser Tyr Leu Gly T/r Asp Cys Thr Thr Asp Lys Cys Asn Asp Arg Gly 610 " 615 620 Val Cys Asn Asn Lys Lys His Cys His Cys Ser Ala Ser Tyr Leu Pro 625 630 635 640 Pro Asp Cys Ser Val Gin Ser Asp Leu Trp Pro Gly Gly Ser He Asp 645 650 655 Ser Gly Asn Phe Pro Pro Val Ala He Pro Ala Arg Leu Pro Glu Arg 660 665 670 Arg Tyr He Glu Asn He Tyr His Ser Lys Pro Met. Arg Trp Pro Phe 675 680 685 Phe Leu Phe He Pro Phe Phe He He Phe Cys Val Leu He Ala He 690 695 700 Met Val Lys Val Asn Phe Gin Arg Lys Lys Trp Arg Thr Glu Asp Tyr 705 710 715 720 Ser Ser Asp Glu Gin Pro Glu Ser Glu Ser Glu Pro Lys Gly 725 730

Claims: WHAT IS CLAIMED IS: 1. A sperm protein in substantially pure form selected from a human PH30 beta chain protein, a mouse PH30 beta chain protein or an amino acid sequence substantially homologous to either the human or mouse PH30 beta chain protein. 2. The sperm protein of Claim 1 , having an integrin binding sequence which is not TDE. 3. The sperm protein of Claim 2, wherein the integrin binding sequence is selected from FEE or QDE. 4. The sperm protein of Claim 1 which is the human PH30 beta chain protein. 5. The sperm protein of Claim 4, having an integrin binding sequence which is FEE. 6. A DNA sequence which encodes the sperm protein of Claim 1 or a portion of the sperm protein sufficient to constitute at least one epitope. 7. The DNA sequence of Claim 6, wherein the epitope is on the native protein. 8. The DNA sequence of Claim 6 which encodes all or a portion of human PH30 beta chain protein. 9. The DNA sequence of Claim 8, wherein the DNA encoding all or a portion of the human PH30 beta protein is characterized by the ability to hybridize, under standard conditions, to the DNA sequence shown in SEQ ID NO: 1. 10. A contraceptive composition comprising a therapeutically effective amount of the protein of Claim 1 , or a polypeptide having the substantially same amino acid sequence as a segment of the protein provided that the polypeptide is sufficient to constitute at least one epitope, and a pharmaceutically acceptable carrier. 11. The contraceptive composition of Claim 10, wherein the epitope is on the native protein. 12. The contraceptive composition of Claim 10, wherein the protein is the human PH30 beta chain protein. 13. The contraceptive composition of Claim 10, wherein the protein is produced by expressing the gene encoding an immunogenic epitope of the sperm protein in a recombinant DNA expression vector. 14. A vector comprising an inserted DNA sequence encoding for the protein of Claim 1. 15. The vector of Claim 14, wherein the inserted DNA sequence is characterized by the ability to hybridize, under standard conditions, to a DNA sequence selected from the DNA sequences of SEQ ID NO: 1 or SEQ ID NO: 3. 16. A host that is compatible with and contains the vector of Claim 14. 17. A method of producing a human or mouse PH30 beta chain sperm protein, comprising the steps of culturing cells containing the DNA of Claim 6 and recovering the sperm protein from the cell culture. 18. The method of Claim 17, wherein the DNA encoding all or a portion of the PH30 beta chain protein is characterized by the ability to hybridize, under standard conditions, to a DNA sequence selected from the DNA sequences of SEQ ID NO: 1 or SEQ ID NO: 3. 19. A method of contraception in a human or mouse subject in need thereof, comprising administering to the subject an amount of the sperm protein of Claim 1 which is effective for the stimulation of antibodies which bind to the sperm protein in vivo. 20. The method of Claim 19, wherein the sperm protein has an integrin binding sequence which is not TDE. 21. A PH30 beta chain protein made by the process of Claim 17. 22. A DNA sequence as shown in Seq. ID No. 1 encoding human PH30 beta chain protein. 23. A purified and isolated DNA sequence consisting essentially of a DNA sequence encoding a polypeptide having an amino acid sequence sufficiently duplicative of that of human or mouse PH30 beta to allow the possession of the biological property of initiating sperm-egg binding or promoting sperm-egg fusion. 24. The DNA sequence of Claim 23 wherein the amino acid sequence contains an integrin binding sequence which is not TDE.

Inventors: ALVES KENNETH, GUPTA SUNIL K, HOLLIS GREGORY FRANKLIN, ALVES, KENNETH, GUPTA, SUNIL, K., HOLLIS, GREGORY, FRANKLIN
Applicants: ALVES KENNETH, GUPTA SUNIL K, HOLLIS GREGORY FRANKLIN, MERCK & CO INC, ALVES, KENNETH, GUPTA, SUNIL, K., HOLLIS, GREGORY, FRANKLIN, MERCK & CO., INC.
Application year: 1995
Main classification: A61K39
IPC codes: A61K38, A61K39, A61P15, C07H21, C07K14, C12N1, C12N5, C12N15, C12P21, C12R1