AU1834401A - OB protein receptor and related compositions and methods - Google Patents

Description

P/00/011 28/5/91 Regulation 3.2

AUSTRALIA

Patents Act 1990

ORIGINAL

COMPLETE SPECIFICATION STANDARD PATENT Name of Applicant: Actual Inventors Address for service is: Amgen Inc.

Ming-Shi CHANG Andrew Avery WELCHER Frederick Addison FLETCHER WRAY ASSOCIATES 239 Adelaide Terrace Perth, WA 6000 Attorney code: WR Invention Title: "OB Protein Receptor and Related Compositions and Methods" This application is a divisional application by virtue of Section 39 of Australian Patent Application 15262/97.

The following statement is a full description of this invention, including the best method of performing it known to me:- 1/2- OB PROTEIN RECEPTOR AND RELATED COMPOSITIONS AND METHODS FIELD OF THE INVENTION The present invention relates to OB protein receptors, related compositions and methods of making and using such receptors and related compositions.

BACKGROUND

Although the molecular basis for obesity is largely unknown, the identification of the "OB gene" and protein encoded ("OB protein") has shed some light on mechanisms the body uses to regulate body fat deposition. Zhang et al., Nature 312: 425-432 (1994); see the Correction at Nature 374: 479 (1995) The OB protein is active in vivo in both ob/ob mutant mice (mice obese due to a defect in the production of the OB gene product) as well as in normal, wild type mice. The biological activity manifests itself in, among other 20 things, weight loss. Siee l, Barinaga, "Obese" Protein Slims Mice, Science 2L2: 475-476 (1995) See PCT International Publication Number WO 96/05309, ."Modulators of Body Weight, Corresponding Nucleic Acids and Proteins, and Diagnostic and Therapeutic Uses Thereof," herein incorporated by reference.

The other biological effects of OB protein are not well characterized. It is known, for instance, that in ob/ob mutant mice, administration of OB protein results in a decrease in serum insulin levels, and serum glucose levels. It is also known that administration of OB protein results in a decrease in body fat. This was observed in both ob/ob mutant mice, as well as non-obese normal mice. Pelleymounter et al., Science 29: 540-543 (1995); Halaas et al., Science 2-ka: 543-546 (1995). See 2 also, Campfield et al., Science 269: 546-549 (1995) (Peripheral and central administration of microgram doses of OB protein reduced food intake and body weight of ob/ob and diet-induced obese mice but not in db/db obese mice.) In none of these reports have toxicities been observed, even at the highest doses.

Despite the promise of clinical application of the OB protein, the mode of action of the OB protein in vivo is not clearly elucidated, in part due to the absence of information on the OB receptor. High affinity binding of the OB protein has been detected in the rat hypothalamus, reportedly indicating OB receptor location. Stephens et al., Nature 377: 530-532 (1995). The db/db mouse displays the identical phenotype as the o 15 ob/ob mouse, extreme obesity and Type II diabetes; this phenotype is thought to be due to a defective OB receptor, particularly since db/db mice fail to respond to OB protein administration. 3ee Stephens et al., 20 Identification of the OB protein receptor is key in determining the pathway of signal transduction.

Moreover, identification of the OB protein receptor would provide powerful application in diagnostic uses, for example, to determine if individuals would benefit from OB protein therapy. Furthermore, the OB receptor could be a key component in an assay for determining additional molecules which bind to the receptor and result in desired biological activity. Further, such soluble receptor could enhance or alter the effectiveness of OB protein (or analog or derivative thereof).

SUMMARY OF THE INVENTION The present invention relates to a novel class of protein receptors, herein denominated "OB protein receptors" or "OB receptors", which are thought to selectively bind OB protein. As such, the novel OB 3 receptor family is provided, as well as novel members of such family. Also provided are nucleic acids, vectors and host cells containing such nucleic acids, related antisense nucleic acids, molecules which selectively bind to the OB protein receptor, and related compositions of matter, such as OB receptor protein/OB protein complexes. In other aspects, the present invention relates to methods of using the above compositions, such as therapeutic and/or diagnostic methods, and methods for preparing OB receptor ligands.

DETAILED DESCRIPTION A novel family of OB receptors is provided.

This novel family resulted from identification of a PCR 15 fragment isolated from a human liver cell cDNA library.

The original PCR fragment, from which primers were isolated, contained a "WSXWS" motif, common to cytokine receptors. As illustrated by the working examples below, using this fragment four members of this OB protein receptor family have been identified. These members, herein designated as and and "D" are indentical at amino acid position 1-891 (using the numbering of Seq. ID No. but diverge at position 892 through the C-terminus. They vary in length at the C-terminus beyond amino acid 891, and the different forms appear to have different tissue distribution.

Using hydrophobicity analysis, the leader sequence is likely to comprise amino acids (Seq. ID.

No. 1) 1-21, 1-22, or 1-28. The first amino acid of the mature protein is likely to be 22 23 or 29 Most likely, based on analysis of eucaryotic cell expression (CHO cell expression see Example 8, infra), the first amino acid of the mature protein is 22(F).

The beginning of the transmembrane domain appears to be located at position 840 or 842 The end of the transmembrane domain appears to be located at position 4 862 863 or 864 Thus, based on predictions from hydrophobicity analysis, for OB protein binding, at a minimum what is needed is the extracellular domain of the mature protein, amino acids 22, 23 or 29 through amino acids 839 or 841 Therefore, the present class of OB receptor proteins includes those having amino acids (according to Seq. ID No. 1): 1-896; 22-896; 23-896; 29-896; 1-839; 22-839; 1-841; 22-841; 23-841; 29-841; 1-891; 22-891; 23-891; 29-891; the amino acids of subparts (1) through having the C-terminal amino acids selected from among: OB receptor B (Seq. ID No. 3) positions 892-904; (ii) OB receptor C (Seq. ID No. positions 892- 958; and, (iii) OB receptor D (Seq. ID No. 7) positions 892-1165; amino acids of subparts b, c, d, f, g, i, j, k, m, n, o, and any of lacking a leader sequence, which have an N-terminal methionyl residue.

Also provided herein is what is thought to be a human splice variant of a soluble OB receptor. This 5 splice variant includes the extracellular domain at least up to amino acid 798 (of Seq. ID No. 1, for example) and has a unique 6 amino acid C-terminus at positions 799-804: G K F T I L.

The functional domains of the OB receptor may be predicted using the information contained in Bazan et al., PNAS-USA 87: 6934-6938 (1990) (incorporated herein by reference). For the present OB receptor, there are two hematopoietin domains, a random coil region, the transmembrane domain, and the intracellular domain. The overall geography may be illustrated as follows: HI1 H2 TM

IC

supr, the domains may be predicted, with essentially an error of approximately plus or minus three base pairs (as applied to all amino acid location specified for purposes of identifying the Bazan predicted domains) The precise locations may be determined empirically by 20 methods known in the art, such as preparing and expressing modified recombinant DNAs. The structural characteristics are though to be important for maintaining the structural integrity of the molecule, and therefore, to the extent that such structure is important for function, for functional characteristics as well.

The hematopoietin domains (HI and H2) are thought to have two fibronectin ty pe 3 repeats each, one set of paired cysteine residues each (thought to form a disulfide bridge), and one "WSXWS box" (referring to the disulfide bridge), and one "WSXWS box" (referring to the single letter amino acid abbreviation, with being any amino acid). The fibrinectin type 3 domains may be identified by location of a double proline which marks the beginning of the second fibronectin type 3 repeat; the actual beginning of such second fibronectin type 3 repeat is likely to begin about 3 amino acids upstream of that double proline.

The first hematopoietin domain is likely to begin at amino acid 123 (using the numbering according to Seq. ID No. 1, for example), which is an isoleucine residue The last amino acid of the hematopoietin domain is likely to be amino acid 339, which is a lysine residue. The two fibronectin type 3 repeats are likely to be located at (about) amino acids 123 through 15 235 and 236 through 339. There is a single pair of cysteine residues which likely form a disulfide bridge, located at position 131 and position 142. The "WSXWS box" is located at position 319 through 323.

The second hematopoietin domain is likely to 20 begin at position 428, which is an isoleucine and end at position 642 which is a glycine The paired fibronectin type 3 repeats are located at about position 428 through position 535 and about position 536 through S. about position 642. One pair of cysteines is located at position 436 and position 447, and the second pair is located at position 473 and 488. The "WSXWS box" is located at position 622-626.

Between the first and the second hematopoietin domain (amino acids 339-428, approximately) is a region of unknown functional significance.

The random coil domain between the H2 and the transmembrane domain, is likely to begin at the amino acid following the end of the second hematopoietin domain, and is likely to end at the beginning of the transmembrane domain. This is likely to be from about amino acid 642 through amino acid 839 7 or 841 (with the transmembrane domain beginning at position 840 or 842 The intracellular domain is likely to begin at position 861 862 863 or 864 The intracellular domain contains three regions, or "boxes," thought to participate in signal transduction (two "JAK" boxes and a single "STAT" box, "Box "Box and "Box With respect to the numbering of the amino acid positions of the form of the OB receptor (Seq. ID No.7, below), box 1 is located at amino acid 871 through 878 Box 2 is located at approximately amino acid number 921 through 931 Box 3 on the form is located at approximately position 1141 through 1144 (amino acids YMPQ, as 15 the "STAT" box is typically a conserved region of "YXXQ" wherein designates any amino acid). The intracellular domain is thought to be responsible for signal transduction. One possible mode of action is via phosphorylation of various residues. See Ihle et al., 20 Cell 84: 331-334 (1996) (Review article, herein incorporated by reference.) One possible mode of action is that upon ligand binding (here, OB protein binding), the OB receptor dimerizes with another receptor. A kinase binds to box 1, and becomes phosphorylated.

(The JAK may already be bound prior to dimerization.) Also, "STATS" bind to box 3 and become phosphorylated on a specific tyrosine. It is thought that this phosphorylation results, probably indirectly, in DNA binding protein production, which results in altered DNA transcription, and therefore altered expression. As seen below in Example 6, one measurement of the capability of an OB receptor to transduce signal is the degree of phosphorylation of JAK/STAT molecules.

The C-terminus region is intracellular (of cell-bound OB receptor). The differences in the C- 8 terminus among members of the present OB receptor family may result in differences in signal transduction among the species. Thus, the present OB receptors include at least the extracellular domain which is important for OB protein ligand binding. Nucleic acids encoding the present OB receptors, vectors, and host cells are also provided for herein.

The extracellular domain may be modified and still retain the function of ligand binding, particularly by one or more of the following modifications: the random coil domain (as indicated above, occuring downstream of the second hematopoietic domain through the beginning of the transmembrane domain) may be deleted (this may be approximately 0 15 positions 642 through 839 or 841); the "WSXWS" box may be modified by substitution of the first serine with another amino acid, particularly conserved in terms of hydrophobicity and/or charge, such as a glycine; (ii) the last serine may be substituted with another amino 20 acid, such as a threonine; (iii) the first tryptophan may be substituted with another amino acid, for example, a tyrosine.

Human genomic DNA encoding OB receptor protein is also provided herein. The genomic DNA has been localized to human chromosome 1P31, which is believed to correspond to mouse chromosome 4, the location of the mouse db locus.

Tissue distribution analysis demonstrates the presence of OB receptor nucleic acids is fairly ubiquitous, and particularly noted in the liver. It is also observed in the ovary, and heart; and, to a lesser extent, in small intestine, lung, skeletal muscle, kidney, and, to an even lesser extent, spleen, thymus, prostate, testes, placenta and pancreas (Example 2, below). There may also be one or more forms of the OB receptor present in serum, such as soluble OB receptor, 9which may be complexed to one or more forms of the OB protein.

Amino Acid Sequences and Compositions According to the present invention, novel OB protein receptors and DNA sequences coding for all or part of such OB receptors are provided. The present invention provides purified and isolated polypeptide products having part or all of the primary structural conformation continuous sequence of amino acid residues) and one or more of the biological properties immunological properties and in vitro biological activity) and physical properties molecular weight) of naturally-occurring mammalian OB receptor 15 including allelic variants thereof. The term "purified and isolated" herein means substantially free of unwanted substances so that the present polypeptides are useful for an intended purpose. For example, one may have a recombinant human OB receptor substantially free 20 of human proteins or pathological agents. These polypeptides are also characterized by being a product of mammalian cells, or the product of chemical synthetic procedures or of procaryotic or eucaryotic host expression by bacterial, yeast, higher plant, insect and mammalian cells in culture) of exogenous DNA sequences obtained by genomic or cDNA cloning or by gene synthesis. The products of expression in typical yeast Saccharomyces cerevisiae), insect, or procaryote F. coli) host cells are free of association with any mammalian proteins. The products of expression in vertebrate non-human mammalian COS or CHO) and avian) cells are free of association with any human proteins. Depending upon the host employed, and other factors, polypeptides of the invention may be glycosylated with mammalian or other eucaryotic carbohydrates or may be non-glycosylated. One may modify 10 the nucleic acid so that glycosylation sites are included in the resultant polypeptide. One may choose to partially or fully deglycosylate a glycosylated polypeptide. Polypeptides of the invention may also include an initial methionine amino acid residue (at position -1 with respect to the first amino acid residue of the mature polypeptide).

In addition to naturally-occurring allelic forms of OB receptor, the present invention also embraces other OB receptor products such as polypeptide analogs of OB receptor and fragments of OB receptor.

Following the procedures of the above noted published application by Alton et al. (WO 83/04053), one can readily design and manufacture genes coding for microbial expression of polypeptides having primary conformations which differ from that herein specified for in terms of the identity or location of one or more residues substitutions, terminal and intermediate additions and deletions). Alternately, modifications of 20 cDNA and genomic genes may be readily accomplished by well-known site-directed mutagenesis techniques and employed to generate analogs and derivatives of OB S: receptor. Such products would share at least one of the biological properties of mammalian OB receptor but may differ in others. As examples, projected products of the invention include those which are foreshortened by deletions; or those which are more stable to hydrolysis (and, therefore, may have more pronounced or longer lasting effects than naturally-occurring); or which have been altered to delete one or more potential sites for glycosylation (which may result in higher activities for yeast-produced products); or which have one or more cysteine residues deleted or replaced by, alanine or serine residues and are potentially more easily isolated in active form from microbial systems; or which have one or more tyrosine residues 11 replaced by phenylalanine; or have an altered lysine composition (such as those prepared for purposes of derivatization). Included are those polypeptides with amino acid substitutions which are "conservative" according to acidity, charge, hydrophobicity, polarity, size or any other characteristic known to those skilled in the art. See enerally, Creighton, Proteins, W.H.

Freeman and Company, (1984) 498 pp. plus index, passim. One may make changes in selected amino acids so long as such changes preserve the overall folding or activity of the protein, (see Table 1, below). Small amino terminal extensions, such as an amino-terminal methionine residue, a small linker peptide of up to about 20-25 residues, or a small extension that facili- 15 tates purification, such as a poly-histidine tract, an antigenic epitope or a binding domain, may also be present. Sea, in general Ford et al., Protein Expression and Purification 2: 95-107, 1991, which is herein incorporated by reference.

*o **ooO°, *g* o o*o *o*o* 12 Table 1 rnnczprV~t-iV;:, Aminnr Clrhrc:c.~c:~~c Id Q Ic S U Q Basic: arginine lysine histidine Acidic: glutamic acid aspartic acid Polar: glutamine asparaQine Hydrophobic: leucine isoleucine valine Aromatic: phenylalanine tryptophan tyrosine Small: glycine alanine serine threonine methionine

S

S.

S

S

S.

5* Also comprehended are polypeptide fragments duplicating only a part of the continuous amino acid sequence or secondary conformations within OB receptor, which fragments may possess one activity of mammalian (particularly human) OB receptor immunological activity) and not others OB protein binding activity).

Of applicability to OB receptor fragments and polypeptide analogs of the invention are reports of the immunological activity of synthetic peptides which substantially duplicate the amino acid sequence extant in naturally-occurring proteins, glycoproteins and nucleoproteins. More specifically, relatively low 13 molecular weight polypeptides have been shown to participate in immune reactions which are similar in duration and extent to the immune reactions of physiologically significant proteins such as viral antigens, polypeptide hormones., and the like. Included among the immune reactions of such polypeptides is the provocation of the formation of specific antibodies in immunologically active animals. See, a Lerner et al., Cell 21: 309-310 (1891); Ross et al., Nature 294: 654-656 (1891); Walter et al., PNAS-USA 77: 5197-5200 (1980); Lerner et al., PNAS-USA, 28: 3403-3407 (1891); Walter et al., PNAS-USA 28: 4882-4886 (1891); Wong et al., PNAS-USA 79: 5322-5326 (1982); Baron et al., Cell o 395-404 (1982); Dressman et al., Nature 295: 185-160 (1982); and Lerner, Scientific American 248: 66-74 (1983). Saa, also, Kaiser et al. Science 223: 249-255 (1984) relating to biological and immunological activities of synthetic peptides which approximately share secondary structures of peptide hormones but may not share their primary structural conformation. The present invention also includes that class of polypeptides coded for by portions of the DNA complementary to the protein-coding strand of the human cDNA or genomic DNA sequences of OB receptor "complementary inverted proteins" as described by Tramontano et al.

Nucleic Acid Res. 12: 5049-5059 (1984). Polypeptides or analogs thereof may also contain one or more amino acid analogs, such as peptidomimetics.

Thus, the present class of 0B receptor proteins includes those having amino acids (according to Seq. ID No. 1): 1-896; 22-896; 23-896; 29-896 1-839; 14 22-839; 29-839; 1-841; 22-841; 23-841; 29-841; 1-891; 22-891; 23-891; 29-891; the amino acids of subparts (1) through having the C-terminal amino acid sequence beginning at position 892 of OB receptor B (Seq. ID No. 3) or C (Seq. ID. No. 15 amino acids of subparts b, c, d, f, Sg, i, j, k, m, n, o, and any of lacking a leader sequence, which have an N-terminal methionyl residue.

Also provided is a longer form of an OB receptor protein, herein denominated the form, which has an amino acid sequence selected from among (according to Seq. ID No. 7): amino acids 1-1165; amino acids 22-1165; amino acids 23-1165; amino acids 29-1165; amino acids of subparts or .0 having an N-terminal methionyl residue.

As set forth above, one may prepare soluble receptor by elimination of the transmembrane and intracellular regions. Examples of soluble receptors include those set forth in Seq. ID Nos. 10 and 13. What is thought to be a native, secreted form of a soluble human OB receptor is also provided herein. This form of OB receptor protein has an amino acid sequence selected from among (according to Seq. ID No. 13): amino acids 1-804; 15 amino acids 22-804; amino acids 23-804; amino acids 29-804; and, amino acids of subparts or having an N-terminal methionyl residue.

In addition, since the C-terminus region of the above polyeptides diverges at position 892 (with respect to Seq. ID Nos. 1, 3, 5, 7 and 13) one may desire to prepare only the polypeptides which are divergent: those having only amino acids 892-896 of Seq. ID No. 1; those having only amino acids 892-904 of Seq. ID No. 3; 15 those having only amino acids 892-958 of Seq. ID No. those having only amino acids 892- 1165 of Seq. ID No. 7; and, those having only amino acids 799-804 20 of Seq. ID No. 13.

The above polypeptides which have an extracellular domain may be modified, as indicated above, and still retain the function of ligand binding.

Such modification may include one or more of the following: the random coil domain (as indicated above, occuring downstream of the second hematopoietic domain through the beginning of the transmembrane domain) may be deleted (this may be approximately positions 642 through 839 or 841); the "WSXWS" box may be modified by substitution of the first serine with another amino acid, particularly conserved in terms of hydrophobicity and/or charge, such as a glycine; (ii) the last serine may be substituted with another amino acid, such as a threonine; (iii) the first tryptophan may be 16 substituted with another amino acid, for example, a tyrosine.

Thus, the present polypeptides include (according to the numbering of Seq. ID No. 7): 1-896; 22-896; 23-896; 29-896 1-839; 22-839; 29-839; 1-841; 22-841; 23-841; 15 29-841; 1-891; 22-891; 23-891; 29-891; the amino acids of subparts (1) o. through having the C-terminal amino acids selected from the C-terminal amino acids of OB receptor B (Seq.

ID No. C (Seq. ID. No. 5) and D (Seq ID No. 7); the amino acids (according to Seq. ID No. 13) selected from the group consisting of 22-804; 23-804 and 29-804; amino acids of subparts b, c, d, f, g, i, j, k, m, n, o, any of lacking a leader sequence, and which have an N-terminal methionyl residue; and amino acids of subparts through which above having at least one of the following modifications: for amino acids of subparts (a) through and those of subpart which are not amino acids according to subpart deletion of (or 17 substitution of amino acid(s) or other modifications of) a random coil domain sequence selected from 640 through 839 (using the numbering according to Seq. ID No. 1); 641 through 839; 642 through 839; 640 through 841; 641 through 841; and 642 through 841; (ii) for amino acids of subpart (q) and those of subpart which contain the sequence of subpart deletion of of (or substitution of amino acid(s) or other modifications of) a random coil domain sequence selected from among: 15 640 through 804; 641 through 804; and, 642 through 804; and, (iii) modification of a "WSXWS" sequence which is substitution of the first serine with another amino acid, particularly conserved in terms of hydrophobicity and/or charge, such as a glycine; substition of the last serine with another amino acid, such as a threonine; and substitution of the first tryptophan with another amino acid, for example, a tyrosine.

One may modify the OB receptor to create a fusion molecule with other peptide sequence. For example, if one desired to "tag" the OB receptor with an immunogenic peptide, one could construct a DNA which would result in such fusion protein. The tag may be at the N-terminus. Also, since it is apparent that the 18 C-terminus is not necessary for ligand binding activity, one may chemically modify the C-terminus of, for example, a soluble OB receptor. One may desire, for example, a preparation whereby one or more polymer molecules such as polyethylene glycol molecules are attached. Thus, another aspect of the present invention is chemically modified OB receptor protein (also further described infra).

An example of such "tag" is provided herein using the C-terminus of a recombinant soluble OB receptor. Seq. ID No. 12 provides a "FLAG-tag" version of such soluble OB receptor (the nucleic acid sequence is provided, which may be transcribed to prepare the polypeptide) Such "FLAG-tag" may also be attached to 15 the N-terminus or other region of an OB receptor protein. This type of "tagging" is useful to bind the protein using reagents, such as antibodies, which are selective for such tag. Such binding may be for detection of the location or amount of protein, or for protein capturing processes where, for example, an affinity column is used to bind the tag, and thus the desired protein. Other types of detectable labels, such as radioisotopes, light-emitting fluorescent or phosporescent compounds), enzymatically cleavable, detectable antibody (or modification thereof), or other substances may be used for such labelling of the present proteins. Detecting protein via use of the labels may be useful for identifying the presence or amount of OB receptor protein or a compound containing such protein OB protein complexed to OB receptor). Moreover, such labelled protein may be useful for distinguishing exogenous OB receptor protein from the endogenous form.

19 Nucleic Acids Novel nucleic acid sequences of the invention include sequences useful in securing expression in procaryotic or eucaryotic host cells of polypeptide products having at least a part of the primary structural conformation and one or more of the biological properties of recombinant human OB receptor. The nucleic acids may be purified and isolated, so that the desired coding region is useful to produce the present polypeptides, for example, or for diagnostic purposes, as described more fully below. DNA sequences of the invention specifically comprise: any of the DNA sequences set forth in Seq. ID No. 2, 4, 6, 8, 9, 11, •15 12, and 14 (and complementary strands); a DNA sequence.which hybridizes (under hybridization conditions disclosed in the cDNA library screening section below, using the 300 bp PCR fragment as described to selectively hybridize to a cDNA encoding an OB receptor protein in a human liver cDNA library, or equivalent conditions or more stringent conditions) to the DNA sequence in subpart or to fragments thereof; and a DNA sequence which, but for the degeneracy of the genetic code, would hybridize to the DNA sequence in subpart Specifically comprehended in parts and are genomic DNA sequences encoding allelic variant forms of human OB receptor and/or encoding OB receptor from other mammalian species, and manufactured DNA sequences encoding OB receptor, fragments of OB receptor, and analogs of OB receptor which DNA sequences may incorporate codons facilitating transcription and translation of messenger RNA in microbial hosts. Such manufactured sequences may readily be constructed according to the methods of Alton et al., PCT published application WO 83/04053.

20 Genomic DNA, such as that of Seq. ID No. 9, encoding the present OB receptors may contain additional non-coding bases, or introns, and such genomic DNAs are obtainable by hybridizing all or part of the cDNA, illustrated in Seq. ID Nos. 2, 4, 6, 8, 11, and 14 to a genomic DNA source, such as a human genomic DNA library.

Such genomic DNA will encode functional OB receptor polypeptide; however, use of the cDNAs may be more practicable in that, since only the coding region is involved, recombinant manipulation is facilitated. The intron/exon location of genomic DNA is set forth in Seq.

ID No. 9, infra.

Nucleic acid sequences include the incorporation of codons which enhance expression by :15 selected nonmammalian hosts; the provision of sites for cleavage by restriction endonuclease enzymes; and the provision of additional initial, terminal or intermediate DNA sequences which facilitate construction of cloning and/or expression vectors.

The present invention also provides DNA sequences coding for polypeptide analogs or derivatives of OB receptor which differ from naturally-occurring forms in terms as described above. The leader sequence DNA may be substituted with another leader sequence for 25 ease in expression or for other purposes.

Also, one may prepare antisense nucleic acids against the present DNAs. Such antisense nucleic acids may be useful in modulating the effects of OB receptor protein in vivo. For example, one may prepare an antisense nucleic acid which effectively disables the ability of a cell to produce OB receptor by binding to the nucleic acid which encodes such OB receptor.

DNA sequences of the invention are also suitable materials for use as labeled probes in isolating human genomic DNA encoding OB receptor, as mentioned above, and related proteins as well as cDNA 21 and genomic DNA sequences of other mammalian species.

DNA sequences may also be useful in various alternative methods of protein synthesis in insect cells) or, as described infra, in genetic therapy in humans and other mammals. DNA sequences of the invention are expected to be useful in developing transgenic mammalian species which may serve as eucaryotic "hosts" for production of OB receptor and OB receptor products in quantity. See, generally, Palmiter et al., Science 222: 809-814 (1983).

Vectors and Host Cells According to another aspect of the present invention, the DNA sequences described herein which encode OB receptor polypeptides are valuable for the information which they provide concerning the amino acid "sequence of the mammalian protein which have heretofore been unavailable. Put another way, DNA sequences provided by the invention are useful in generating new e and useful viral and circular plasmid DNA vectors, new and useful transformed and transfected procaryotic and eucaryotic host cells (including bacterial cells, yeast cells, insect cells, and mammalian cells grown in culture), and new and useful methods for cultured growth of such host cells capable of expression of OB receptor and its related products.

The DNA provided herein (or corresponding RNAs) may also be used for gene therapy for, example, treatment of conditions characterized by the overexpression of OB protein, such as anorexia or cachexia. Alternatively, gene therapy may be used in cases where increased sensitivity to OB protein is desired, such as in cases where an individual has a condition characterized by OB protein receptors defective in ability to bind or retain the binding of OB protein. Currently, vectors suitable for gene therapy 22 (such as retroviral or adenoviral vectors modified for gene therapy purposes and of purity and pharmaceutical acceptability) may be administered for delivery into the lung, for example. Such vectors may incorporate nucleic acid encoding the present polypeptides for expression in a desired location. Gene therapy may involve more than one gene for a desired protein or different desired proteins.

Alternatively, one may use no vector so as to facilitate relatively stable presence in the host. For example, homologous recombination of a DNA as provided herein or of a suitable transcription or translation control region may facilitate integration into or expression from a host genome. (This may be performed 15 for production purposes as well, U.S. Patent No. 5,272,071 and WO 91/09955.) The nucleic acid may be placed within a pharmaceutically acceptable carrier to facilitate cellular uptake, such as a lipid solution carrier a charged lipid), a liposome, or polypeptide carrier polylysine). A review article on gene therapy is Verma, Scientific American, November 1990, pages 68-84 which is herein incorporated by reference.

Thus, the present invention provides for a population of cells expressing an OB receptor of the present OB receptor family. Such cells are suitable for transplantation or implantation into an individual for therapeutic purposes. For example, one may prepare a population of cells to overexpress OB receptor (such as one identified in the. Sequence ID's or otherwise denoted herein), or to express a desired form of OB receptor, such as one which is particularly sensitive to OB protein a form which has a desired capacity for signal transduction). One may then implant such cells into an individual to increase that individual's sensitivity to OB protein. Such cells may, for example, 23 be liver cells, bone marrow cells, or cells derived from umbillical cord. Alternatively, one may wish to use overexpressing circulating cells such as blood progenitor cells, T cells or other blood cells. For humans, human cells may be used. Cells may be in the form of tissue. Such cells may be cultured prior to transplantation or implantation. Such OB receptor overexpression, or expression of particularly sensitive forms of OB receptor may be accomplished by, for example, altering the regulatory mechanism for expression of OB receptor, such as using homologous recombination techniques as described supra. Thus, provided is a population of host cells modified so that expression of endogenous OB receptor DNA is enhanced.

15 The cells to be transferred to the recipient may be cultured using one or more factors affecting the growth or proliferation of such cells if appropriate.

Hematopoietic factors may be used in culturing hematopoietic cells. Such factors include G-CSF, EPO, MGDF, SCF, Flt-3 ligand, interleukins IL1-IL13), GM-CSF, LIF, and analogs and derivatives thereof as available to one skilled in the art.

Nerve cells., such as neurons or glia, may also be used, and these may be cultured with neurotrophic 25 factors such as BDNF, CNTF, GDNF, NT3, or others.

There may be a co-gene therapy involving the transplantation of cells expressing more than one desired protein. For example, cells expressing OB receptor protein may be used in conjunction, simultaneously or in serriatim with cells expressing OB protein.

For gene therapy dosages, one will generally use between one copy and several thousand copies of the present nucleic acid per cell, depending on the vector, the expression system, the age, weight and condition of the recipient and other factors which will be apparent 24 to those skilled in the art. The cellular delivery of such protein may be designed to last for a selected period of time, such as a period of days, weeks, months or years. At the end of the effective time period, the recipient of such transformed cells may receive another "dose" transplantation of cells). Cells may be selected for their lifespan, their time period of expression of the desired protein, or their ability to be reisolated from an individual for blood cells, leukaphoresis may be used to retrieve transformed cells using markers present on the cell surface). Vectors may be similiarly designed using, for example, viruses which have a known period of expression of DNAs contained therein.

The desired cells or vectors may be stored using techniques, such as freezing, available to those in the art.

Thus, the present invention also contemplates a method for administering OB: receptor protein to an individual, wherein the source of said OB receptor protein is selected from a population of cells expressing OB receptor protein and (ii) a population of vectors expressing OB receptor protein. Said OB receptor protein may be selected from among those described herein. Said vectors may be virus vectors capable of infecting human cells. Said cells may be selected from among tissue or individual cells. Said individual cells may be selected from among adipocytes, fibroblasts, bone marrow cells, peripheral blood progenitor cells, red blood cells, and white blood cells, including T cells and nerve cells. Said population of cells or vectors may be co-administered with a population of cells or vectors which express OB protein or another desired protein. Said cells or vectors may be stored for use in an individual. Storage may be by freezing 25 Complexes In addition to the OB receptor protein as described herein, one may prepare complexes of OB receptor protein and OB protein, analog or derivative.

The OB protein may be selected from those described in PCT publication WO 96/05309, above and hereby incorporated by reference in its entirety.

Figure 3 of that publication (Seq. ID No. 4, as cited therein) depicts the full deduced amino acid sequence derived for the human OB gene. The amino acids are numbered from 1 to 167. A signal sequence cleavage site is located after amino acid 21 (Ala) so that the mature protein extends from amino acid 22 (Val) to amino acid 167 (Cys). For the present disclosure, a different numbering, is used herein, where the amino acid position 1 is the Valine residue which is at the beginning of the mature protein.

Generally, the OB protein for use will be capable of complexing to the OB protein receptor selected. Thus, one may empirically test the binding capability (to all or part of the extracellular domain of the OB receptor as indicated above) to determine which OB protein forms may be used. Generally, modifications generally applicable as indicated above for OB receptor protein may also be applied here, and that disclosure is incorporated by reference here. As set forth in WO 96 05309, OB protein in its native form, or fragments (such as enzyme cleavage products) or other truncated forms, analogs, and derivatives all retain biological activity. Such forms may be used so long as the form binds to at least a portion of the extracellular domain of the present OB receptor proteins.

An effective amount of an OB protein, analog or derivative thereof may be selected from among 26 according to the amino acid sequence as presented in PCT WO 96/05309, Figure 3 numbered so that the first amino acid of the mature protein is number 1: the amino acid sequence 1-146, optionally lacking a glutaminyl residue at position 28, and further optionally having a methionyl residue at the N-terminus; an amino acid sequence of subpart having a different amino acid substituted in one or more of the following positions: 4, 8, 32, 33, 35, 48, 53, 60, 64, 66, 67, 68, 71, 74, 77, 78, 89, 97, 100, 102, 105, 106, 107, 108, 111, 112, 118, 136, 138, 142, and 145; a truncated OB protein analog 15 selected from among: (using the numbering of subpart (a) above): amino acids 98-146 (ii) amino acids 1-32 (iii) amino acids 1-35 20 (iv) amino acids 40-116 amino acids 1-99 and 112-146 (vi) amino acids 1-99 and 112-146 having one or more of amino acids 100-111 sequentially placed between amino acids 99 and 112; and, (vii) the truncated OB analog of subpart having one or more of amino acids 100, 102, 105, 106, 107, 108, 111, 112, 118, 136, 138, 142, and 145 substituted with another amino acid; (viii) the truncated analog of subpart (ii) having one or more of amino acids 4, 8 and 32 substituted with another amino acid; (ix) the truncated analog of subpart (iv) having one or more of amino acids 50, 53, 64, 66, 67, 68, 71, 74, 77, 78, 89, 97, 100, 102, 27 105, 106, 107, 108, 111 and 112 replaced with another amino acid; the truncated analog of subpart having one or more of amino acids 4, 8, 32, 33, 35, 48, 50, 53, 60, 64, 66, 67, 68, 71, 74, 77, 78, 89, 97, 112, 118, 136, 138, 142, and 145 replaced with another amino acid; (xi) the truncated analog of subpart (vi) having one or more of amino acids 4, 8,32, 33, 35, 48, 50, 53, 60, 64, 66, 67, 68, 71, 74, 77, 78, 89, 97, 100, 102, 105, 106, 107, 108, 111, 112, 118, 136, 138, 142, and 145 replaced with another amino acid; (xii) the truncated analog of any of 15 subparts having an N-terminal methionyl residue; and the OB protein or analog derivative of any of subparts through comprised of a chemical moiety connected to the protein moiety; a derivative of subpart wherein said chemical moiety is a water soluble polymer moiety; a derivative of subpart wherein said water soluble polymer moiety is polyethylene glycol; 25 A derivative of subpart wherein said water soluble polymer moiety is a polyamino acid moiety; a derivative of subpart wherein said water soluble polymer moiety is attached at solely the N-terminus of said protein moiety; an OB protein, analog or derivative of any of subparts through in a pharmaceutically acceptable carrier.

OB proteins, analogs and related molecules are also reported in the following publications; however, no 28 representation is made with regard to the activity of any composition reported: U.S.Patent Nos. 5,521,283; 5,532,336; 5,552,522; 5,552,523; 5,552,524; 5,554,727; 5,559,208; 5,563,243; 5,563,244; 5,563,245; 5,567,678; 5,567,803; 5,569,744; 5,569,743 (all assigned to Eli Lilly and Company); PCT W096/23517; W096/23515; W096/23514; W096/24670; W096/23513; W096/23516; W096/23518; W096/23519; W096/23520; W096/23815; W096/24670; W096/27385 (all assigned to Eli Lilly and Company); PCT W096/22308 (assigned to Zymogenetics); PCT W096/29405 (assigned to Ligand 15 Pharmaceuticals, Inc.); PCT W096/31526 (assigned to Amyin "Pharmaceuticals, Inc.); PCT W096/34885 (assigned to Smithkline Beecham PLC) PCT W096/35787 (assigned to Chiron); EP 0 725 079 (assigned to Eli Lilly and o**o Company); EP 0 725 078 (assigned to Eli Lilly and Company); EP 0 736 599 (assigned to Takeda); EP 0 741 187 (assigned to F. Hoffman LaRoche).

To the extent these references provide for useful OB proteins or analogs or derivatives thereof, or associated compositions or methods, such compositions and/or methods may be used in conjunction with the present OB receptor proteins, such as for coadministration (together or separately, in a selected dosage schedule) or by complexing compositions to the present OB protein receptors. With the above provisos, these publications are herein incorporated by reference.

29 Derivatives and Formulations The present OB protein receptor and/or OB protein (herein the term "protein" is used to include "peptide" and OB protein or receptor analogs, such as those recited infra, unless otherwise indicated) may also be derivatized by the attachment of one or more chemical moieties to the protein moiety. If the present pharmaceutical compositions contain as the active ingredient a complex of OB protein receptor and OB protein, one or both of such proteins may be derivatized. The chemically modified derivatives may be further formulated for intraarterial, intraperitoneal, intramuscular, subcutaneous, intravenous, oral, nasal, pulmonary, topical or other routes of administration.

Chemical modification of biologically active proteins has been found to provide additional advantages under certain circumstances, such as increasing the stability and circulation time of the therapeutic protein and decreasing immunogenicity. See U.S. Patent No. 4,179,337, Davis et al., issued December 18, 1979.

For a review, see Abuchowski et al., in Enzymes as Drugs. Holcerberg and J. Roberts, eds.

pp. 367-383 (1891)). A review article describing protein modification and fusion proteins is Francis, Focus on Growth Factors 1: 4-10 (May 1992) (published by Mediscript, Mountview Court, Friern Barnet Lane, London OLD, UK).

Preferably, for therapeutic use of the end-product preparation, the chemical moiety for derivatization will be pharmaceutically acceptable. A polymer may be used. One skilled in the art will be able to select the desired polymer based on such considerations as whether the polymer/protein conjugate will be used therapeutically, and if so, the desired dosage, circulation time, resistance to proteolysis, and 30 other considerations. For the present proteins and peptides, the effectiveness of the derivatization may be ascertained by administering the derivative, in the desired form by osmotic pump, or by injection or infusion, or, further formulated for oral, pulmonary or nasal delivery, for example), and observing biological effects as described herein.

The chemical moieties suitable for derivatization may be selected from among various water soluble polymers. The polymer selected should be water soluble so that the protein to which it is attached so that it is miscible in an aqueous environment, such as a physiological environment. The water soluble polymer may be selected from the group consisting of, for 15 example, polyethylene glycol, copolymers of ethylene glycol/propylene glycol, carboxymethylcellulose, dextran, polyvinyl alcohol, polyvinyl pyrolidone, poly-1, 3-dioxolane, poly-1,3,6-trioxane, ethylene/maleic anhydride copblymer, polyaminoacids (either homopolymers or random or non-random copolymers (see supra regarding fusion molecules), and dextran or poly(n-vinyl pyrolidone)polyethylene glycol, propylene glycol homopolymers, polypropylene oxide/ethylene oxide co-polymers, polyoxyethylated polyols, polystyrenemaleate and polyvinyl alcohol. Polyethylene glycol propionaldenhyde may have advantages in manufacturing due to its stability in water.

Fusion proteins may be prepared by attaching polyaminoacids to the OB protein receptor or OB protein (or analog or complex) moiety. For example, the polyamino acid may be a carrier protein which serves to increase the circulation half life of the protein. For the present therapeutic or cosmetic purposes, such polyamino acid should be those which do not create neutralizing antigenic response, or other adverse response. Such polyamino acid may be selected from the 31 group consisting of serum album (such as human serum albumin), an antibody or portion thereof (such as an antibody constant region, sometimes called or other polyamino acids. As indicated below, the location of attachment of the polyamino acid may be at the N-terminus of the OB protein moiety, or other place, and also may be connected by a chemical "linker" moiety to the OB protein.

The polymer may be of any molecular weight, and may be branched or unbranched. For polyethylene glycol, the preferred molecular weight is between about 2 kDa and about 100 kDa (the term "about" indicating that in preparations of polyethylene glycol, some molecules will weigh more, some less, than the stated molecular weight) for ease in handling and manufacturing. Other sizes may be used, depending on the desired therapeutic profile the duration of sustained release desired, the effects, if any on biological activity, the ease in handling, the degree or •20 lack of antigenicity and other known effects of the polyethylene glycol to a therapeutic protein or analog) The number of polymer molecules so attached may vary, and one skilled in the art will be able to ascertain the effect on function. One may monoderivatize, or may provide for a di-, tri-, tetra- or some combination of derivatization, with the same or *0 different chemical moieties polymers, such as different weights of polyethylene glycols). The proportion of polymer molecules to protein (or peptide) molecules will vary, as will their concentrations in the reaction mixture. In general, the optimum ratio (in terms of efficiency of reaction in that there is no excess unreacted protein or polymer) will be determined by factors such as the desired degree of derivatization mono, di-, tri-, etc.), the molecular weight of 32 the polymer selected, whether the polymer is branched or unbranched, and the reaction conditions.

The chemical moieties should be attached to the protein with consideration of effects on functional or antigenic domains of the protein. There are a number of attachment methods available to those skilled in the art. EP 0 401 384 herein incorporated by reference (coupling PEG to G-CSF), see also Malik et al., Exp.

Hematol. 1028-1035 (1992) (reporting pegylation of GM-CSF using tresyl chloride). For example, polyethylene glycol may be covalently bound through amino acid residues via a reactive group, such as, a free amino or carboxyl group. Reactive groups are those to which an activated polyethylene glycol molecule (or 15 other chemical moiety) may be bound. The amino acid residues having a free amino group may include lysine Sresidues and the N-terminal amino acid residue. Those having a free carboxyl group may include aspartic acid residues, glutamic acid residues, and the C-terminal amino acid residue. Sulfhydrl groups may also be used as a reactive group for attaching the polyethylene glycol molecule(s) (or other chemical moiety).

SPreferred for therapeutic manufacturing purposes is .attachment at an amino group, such as attachment at the N-terminus or lysine group. Attachment at residues important for receptor binding should be avoided if receptor binding is desired.

One may specifically desire N-terminally chemically modified protein. Using polyethylene glycol as an illustration of the present compositions, one may select from a variety of polyethylene glycol molecules (by molecular weight, branching, etc.), the proportion of polyethylene glycol molecules to protein molecules in the reaction mix, the type of pegylation reaction to be performed, and the method of obtaining the selected N-terminally pegylated protein. The method of obtaining 33 the N-terminally pegylated preparation separating this moiety from other monopegylated moieties if necessary) may be by purification of the N-terminally pegylated material from a population of pegylated protein molecules. Selective N-terminal chemical modification may be accomplished by reductive alkylation which exploits differential reactivity of different types of primary amino groups (lysine versus the N-terminal) available for derivatization in a particular protein. See PCT WO 96/11953, herein incorporated by reference. Under the appropriate reaction conditions, substantially selective derivatization of the protein at the N-terminus with a carbonyl group containing polymer is achieved. For example, one may selectively 15 N-terminally pegylate the protein by performing the reaction at a pH which allows one to take advantage of the pKa differences between the e-amino group of the lysine residues and that of the a-amino group of the N-terminal residue of the protein. By such selective 20 derivatization, attachment of a polymer to a protein is controlled: the conjugation with the polymer takes place predominantly at the N-terminus of the protein and no significant modification of other reactive groups, such as the lysine side chain amino groups, occurs.

Using reductive alkylation, the polymer may be of the type described above, and should have a single reactive aldehyde for coupling to the protein. Polyethylene glycol propionaldehyde, containing a single reactive aldehyde, may be used.

An N-terminally chemically modified derivative is preferred (over other forms of chemical modification) for ease in production of a therapeutic. N-terminal chemical modification ensures a homogenous product as characterization of the product is simplified relative to di-, tri- or other multi-derivatized products. The use of the above reductive alkylation process for 34 preparation of an N-terminally chemically modified product is preferred for ease in commercial manufacturing.

In yet another aspect of the present invention, provided are methods of using pharmaceutical compositions of the proteins, and derivatives. Such pharmaceutical compositions may be for administration by injection, or for oral, pulmonary, nasal, transdermal or other forms of administration. In general, comprehended by the invention are pharmaceutical compositions comprising effective amounts of protein or derivative products of the invention together with pharmaceutically acceptable diluents, preservatives, solubilizers, emulsifiers, adjuvants and/or carriers. Such 15 compositions include diluents of various buffer content Tris-HCl, acetate, phosphate), pH and ionic strength; additives such as detergents and solubilizing agents Tween 80, Polysorbate 80), anti-oxidants ascorbic acid, sodium metabisulfite), preservatives Thimersol, benzyl alcohol) and S• bulking substances lactose, mannitol); o incorporation of the material into particulate preparations of polymeric compounds such as polylactic acid, polyglycolic acid, etc. or into liposomes. See, PCT WO96/29989, Collins et al., "Stable protein: phospholipid compositions and methods," published October 3, 1996, herein incorporated by reference.

Hylauronic acid may also be used, and this may have the effect of promoting sustained duration in the circulation. Such compositions may influence the physical state, stability, rate of in vivo release, and rate of in vivo clearance of the present proteins and derivatives. See, e Remington's Pharmaceutical Sciences, 18th Ed. (1990, Mack Publishing Co., Easton, PA 18042) pages 1435-1712 which are herein incorporated by reference. The compositions may be prepared in 35 liquid form, or may be in dried powder, such as lyophilized form. Implantable sustained release formulations are also contemplated, as are transdermal formulations.

Specifically contemplated are oral dosage forms of the above derivatized proteins. Protein may be chemically modified so that oral delivery of the derivative is efficacious. Generally, the chemical modification contemplated is the attachment of at least one moiety to the protein (or peptide) molecule itself, where said moiety permits inhibition of proteolysis; and uptake into the blood stream from the stomach or intestine. Also desired is the increase in overall stability of the protein and increase in circulation time in the body. See PCT W095/21629, Habberfield, "Oral Delivery of Chemically Modified Proteins" (published August 17, 1995) herein incorporated by reference, and U.S. Patent No. 5,574,018, Habberfield et al., "Conjugates of Vitamin B12 and Proteins," issued 20 November 12, 1996, herein incorporated by reference.

Also contemplated herein is pulmonary delivery of the present protein, or derivative thereof. The protein (derivative) is delivered to the lungs of a mammal while inhaling and traverses across the lung epithelial lining to the blood stream. See, PCT W094/20069, Niven et al., "Pulmonary administration of granulocyte colony stimulating factor, published September 15, 1994, herein incorporated by reference.

Nasal delivery of the protein (or analog or derivative) is also contemplated. Nasal delivery allows the passage of the protein to the blood stream directly after administering the therapeutic product to the nose, without the necessity for deposition of the product in the lung. Formulations for nasal delivery include those with absorption enhancing agents, such as dextran or 36 cyclodextran. Delivery via transport across other mucous membranes is also contemplated.

Dosaes One skilled in the art will be able to ascertain effective dosages by administration and observing the desired therapeutic effect. Preferably, the formulation of the molecule or complex in a pharmaceutical composition will be such that between about .10 gg/kg/day and 10 mg/kg/day will yield the desired therapeutic effect. The effective dosages may be determined using diagnostic tools over time. For example, a diagnostic for measuring the amount of OB protein or OB receptor protein in the blood (or plasma 15 or serum) may first be used to determine endogenous levels of OB protein (or receptor). Such diagnostic S• tool may be in the form of an antibody assay, such as an *"antibody sandwich assay. The amount of endogenous OB receptor protein (such as soluble receptor) is quantified initially, and a baseline is determined. The S• therapeutic dosages are determined as the quantification ~of endogenous and exogenous OB receptor protein (that is, protein, analog or derivative found within the body, either self-produced or administered) is continued over the course of therapy. The dosages may therefore vary over the course of therapy, with a relatively high dosage being used initially, until therapeutic benefit is seen, and lower dosages used to maintain the therapeutic benefits.

During an initial course of therapy of an obese person, dosages may be administered whereby weight loss and concomitant fat tissue decrease increase is achieved. Once sufficient weight loss is achieved, a dosage sufficient to prevent re-gaining weight, yet sufficient to maintain desired weight or fat mass may be administered. These dosages can be determined 37 empirically, as the effects of OB protein are reversible. Campfield et al., Science 269: 546-549 (1995) at 547. Thus, if a dosage resulting in weight loss is observed when weight loss is not desired, one would administer a lower dose,. yet maintain the desired weight.

Therapeutic Compositions and Methods The present OB receptor proteins, alone, or in combination with an OB protein, and nucleic acids may be used for methods of treatment, or for methods of manufacturing medicaments for treatment. Such treatment includes conditions characterized by excessive production of OB protein, wherein the present OB receptors, particularly in soluble form, may be used to complex to and therefore inactivate such excessive OB protein. Or, such OB receptor protein, particularly in soluble form, may act to protect the activity of OB protein. While not wishing to be bound by theory, one 20 may postulate that OB protein receptor agonist activity may be accomplished by a protective effect achieved when OB protein receptor (particularly soluble receptor) is complexed to OB protein. Such effect may prolong the serum half life of OB protein in vivo. Such treatments may be accomplished by preparing soluble receptor use of an extracellular domain as described sura) and administering such composition to an individual in need thereof or by preparation of a population of cells containing or expressing such OB receptor, and transplanting such cells into the individual in need thereof.

The present OB receptors may also be used for treatment of those having defective OB receptors. For example, one may treat an individual having defective OB receptors by preparation of a population of cells containing such non-defective OB receptor, and -38 transplanting such cells into an individual. Or, an individual may have an inadequate number of OB receptors, and cells containing such receptors may be transplanted in order to increase the number of OB receptors available to an individual.

The present OB receptor proteins and related compositions such as OB receptor protein/OB protein complex, provide for weight loss, fat loss, increase in lean mass, increase in insulin sensitivity, increase in overall strength, increase in red blood cells (and oxygenation in the blood), decrease in bone resportion or osteoporosis, decreased or maintained serum cholesterol level, decreased or maintained triglyceride (LDL or VLDL) levels, prevention or reduction in arterial plaque formation, treatment of hypertension, and prevention or reduction of gall stone formation. As body fat composition may be correlated with certain types of cancers, the present compositions may be useful e* for the prevention or amelioration of certain types of cancers. The present invention also includes methods for manufacture of a medicament for use in conjunction with the cosmetic/therapeutic conditions described herein, containing at least one of the present compositions.

The present compositions and methods may be used in conjunction with other medicaments, such as those useful for the treatment of diabetes insulin or analogs thereof, thiazolidinediones or other antihyperglycemic agents, and possibly amylin or antagonists there of), cholesterol and blood pressure lowering medicaments (such as those which reduce blood lipid levels or other cardiovascular medicaments), and activity increasing medicaments amphetamines).

Appetite suppressants may also be used (such as serotonin modulators and neuropeptide Y antagonists).

39 Such administration may be simultaneous or may be in seriatim.

In addition, the present methods may be used in conjunction with surgical procedures, such as cosmetic surgeries designed to alter the overall appearance of a body liposuction or laser surgeries designed to reduce body mass, or implant surgeries designed to increase the appearance of body mass). The health benefits of cardiac surgeries, such as bypass surgeries or other surgeries designed to relieve a deleterious condition caused by blockage of blood vessels by fatty deposits, such as arterial plaque, may be increased with concomitant use of the present compositions and methods. Methods to eliminate 15 gall stones, such as ultrasonic or laser methods, may oo o also be used either prior to, during or after a course of the present therapeutic methods. Furthermore, the present methods may be used as an adjunct to surgeries or therapies for broken bones, damaged muscle, or other 20 therapies which would be improved by an increase in lean tissue mass.

In yet another aspect, the present invention provides for methods of manufacture of a medicament for the treatment of obesity, type II diabetes, excess blood lipid, or cholesterol levels, increasing sensitivity to ".insulin, increasing lean mass, and other conditions as "'.set forth above. Also provided are solely cosmetic treatments for individuals wishing to improve appearance by weight loss, and more specifically, loss of fat deposits, even in the absence of any therapeutic benefit.

Diagnostic Compositions and Methds As indicated supra, polypeptide products of the invention may be "labeled" by association with a detectable marker substance radiolabeled with 125I, fluorescent, chemiluminescent, enzyme) to provide reagents useful in detection and quantification of OB receptor (or complexes) in solid tissue and fluid samples such as blood or urine. Nucleic acid products of the invention may also be labeled with detectable markers (such as radiolabels and non-isotopic labels such as biotin) and employed in hybridization processes to locate the human OB receptor gene position and/or the position of any related gene family in a chromosomal map. Nucleic acid sequences which selectively bind the human OB receptor gene are useful for this purpose.

They may also be used for identifying human OB receptor gene disorders at the DNA level and used as gene markers for identifying neighboring genes and their disorders.

Such nucleic acid sequences may be sued for detection or measurement of OB receptor mRNA level from a biological sample. Contemplated herein are kits containing such labelled materials.

The protein and/or nucleic acids provided herein may also be embodied as part of a kit or article of manufacture. Contemplated is an article of manufacture comprising a packaging material and one or more preparations of the presently provided compositions. Such packaging material will comprise a label indicating that the protein or nucleic acid preparation is useful for detecting and/or quantifying the amount of OB receptor in a biological sample, or OB receptor defects in a biological sample. As such, the kit may optionally include materials to carry out such testing, such as reagents useful for performing DNA or RNA hybridization analysis, or PCR analysis on blood, urine, or tissue samples.

A further embodiment of the invention is selective binding molecules, such as monoclonal antibodies selectively binding GB receptor. The 41 hybridoma technique described originally by Kohler and Milstein Eur. J. Immunol. 6, 511-519 (1976) has been widely applied to produce hybrid cell lines that secrete high levels of monoclonal antibodies against many specific antigens. Recombinant antibodies, (se Huse et al., Science 246: 1275 (1989)) may also be prepared.

Such recombinant antibodies may be further modified, such as by modification of complementarity determining regions to increase or alter affinity, or "humanizing" such antibodies. Such antibodies may be incorporated into a kit for diagnostic purposes, for example. A diagnostic kit may be employed to determine the location and/or amount or OB receptor of an individual.

Diagnostic kits may also be used to determine if an individual has receptors which bind OB protein, or those which, to varying degrees, have reduced binding capacity or ability. As stated infra, such antibodies may be prepared using immunogenic portions of an OB receptor protein. Such selective binding molecules may 20 themselves be alternatives to OB protein, and may be formulated for pharmaceutical composition.

Such proteins and/or nucleic acids may be used for tissue distribution assays (for example, as provided in the working example below) or for other assays to determine the location of OB receptor.

The present OB receptor protein family may be used in methods to obtain OB protein analogs, mimetics or small molecules. One would simply prepare a desired :ee OB receptor protein, particularly one with capability of binding to native OB protein, and assay the test molecule, which may be labelled with a detectable label substance, for ability to bind to such receptor. Other parameters, such as affinity, and location of binding, may also be ascertained by methods available to those skilled in the art. For example, one could use portions of the present OB receptors, particularly portions in 42 the extracellular domain which are necessary for ligand binding, to determine the location of such binding. One could prepare OB receptors which have various truncations or deletions of regions of the extracellular domain which could be used to determine the location of test molecule binding. One could use an OB receptor known to be defective in native OB binding, such as potentially one from an individual having such defective receptors, and use this as the basis for ascertaining OB protein which would be effective to result in desired biological activity weight loss, reduction in blood dyslipidemias or lowering of cholesterol levels, reduction in incidence or severity of diabetes). Other uses include solely cosmetic uses for alteration of body appearance, particularly the removal of fat.

The present OB receptor protein or nucleic acids may also be useful to identify substances which "up-regulate" OB protein or receptor. For instance, the temporal expression of OB receptor in vivo may be useful to determine if an administered substance causes an increase or decrease in OB receptor. One may conclude that an increase in OB receptor expression results in modultion of weight or lipid metabolism.

The divergence in the C-terminus may represent OB receptors with different signal transduction abilities. Therefore the different receptor family members may be used for different assays, depending on the type of signal transduction observed. It is thought that at least a portion of the intracellular domain is necessary for signal transduction (see supra).

The following examples are offered to more fully illustrate the invention, but are not to be construed as limiting the scope thereof.

43 EXAMPLE 1: IDENTIFICATION OF HUMAN OB RECEPTOR PROTEIN Human OB receptor protein DNA was identified in a human liver cDNA library in two steps. The first step used two primers in polymerase chain reaction (PCR) to amplify a selected 300 base pair region from the human liver cDNA library. The second step used the PCR fragment as a probe to screen the human liver cDNA library. Thirteen clones were obtained, but these were incomplete at the 5' end. A procedure was performed to complete the 5' end to make complete clones. Twelve clones were sequenced. These twelve clones were identified as either or as denoted by the C-terminus of the predicted amino acid sequence.

Polymerase Chain Reaction.

The original PCR primer was based on the end and the 3' end of a 416 base pair sequence having 20 GenBank Database Accession No. T73849. This sequence was selected on the basis of a known motif present in cytokine receptors, "WSXWS".

The 5' primer had the sequence 73-96 of the 416 bp sequence. The 3' primer had the sequence 337-360 of the 416 bp sequence.

These primers were used to probe a human cDNA liver library (Stratagene). Standard methods were used.

This resulted in a PCR fragment having the sequence 73-360 of the 416 bp fragment.

Hybridization.

The 300 bp PCR fragment was used to probe a human liver cDNA library (Stratagene) using standard methods. This second hybridization resulted in 13 positive clones. These were partial clones, incomplete at the 5' end.

44 Completion of the 5' end.

Rapid Amplification of cDNA End ("RACE", kit, GIBCO/BRL) was used to obtain the full length clones.

Secuencina results.

Sequencing revealed the three types of OB receptor DNAs. Of the thirteen clones, 4 clones were the type (Seq. ID Nos. 1 and 1 clone was the "B" type (Seq. ID Nos. 3 and 4) and 4 clones were of the "C" type (Seq. ID Nos. 5 and 6).

As can be seen from the Sequence Identifications (below), OB receptor A is 896 amino acids long, is 904 amino acids long, and is 958 amino acids long. These different OB receptors are identical at amino acid positions 1-891, and diverge almost completely beginning at position 892. The leader sequence is postulated to be, by hydrophobicity analysis, amino acids 1-21(M-A), 1-22(M-F) or 1-28(M-I), with the mature protein beginning at positions 22(F), 23(N) or 29(T). Based on hydrophobicity analysis, the leader sequence is most likely to be at positions 1-21(M through Chinese Hamster Ovary Cell cell production of the secreted form of OB receptor protein also produced a protein having amino acid number 22 as the first amino acid of the mature protein. The transmembrane region is likely to begin at either position 840 or 842(L) through position 862(I), 863(S) or 864(H). For OB receptor type the last 30 amino acid is located at position 896 and is a lysine For OB receptor type the last amino acid is located at position 904 and is a glutamine For OB receptor type the last amino acid is located at position 958 and is glutamic acid For OB receptor protein type the Cterminal region possesses high homology to a known human 45 transposable element. From nucleotide 2737 through 2947 of the present human OB receptor protein type there is a 98.1% homology with a 211 base section of a human retrotransposable element described in Ono et al., Nucl.

Acids Res. 15: 8725-8737 (1987) (bases 520 through 731, SINE-R11, GENBANK accession no. x07417).

EXAMPLE 2: TISSUE DISTRIBUTION Tissue distribution was ascertained using two methods. The first method involved using the entire type OB receptor. The second method involved using probes which are specific to the C-terminal region of the protein. Since these C terminal regions are divergent, the second method detected the tissue distribution of the different members of the OB receptor family.

The first method used a Northern Blot kit (Clontech), using the entire type A OB receptor DNA as a probe. The second method used PCR with primers specific 20 to the nucleic acids encoding the divergent C terminus of the three types. Standard methods were used.

Table 2 shows the results for the Northern Blot and the PCR methods. The indicates the investigator's subjective determination of the strength of signal. For the Northern Blot analysis, a triple indicates that a result (a dark "band" on the Xray film) was seen upon overnight exposure of the film.

A double indicates that bands were seen at two weeks of exposure. A single indicates that the 30 bands were seen after three weeks of exposure. In addition, using this method, two molecular weights were observed, one at 4 Kb and one at 6.2 Kb. Although distribution was ubiquitous, the strongest signals were seen for ovary, heart and liver. For the PCR analysis, OB receptor was seen in all tissue types tested (prostate, ovary, small intestine, heart, lung, liver 46 and skeletal muscle), type was seen only in lung and liver, and type was seen in ovary, heart, lung and liver.

Table 2 Tissue Distribution of the Novel OB Receotor Blt 4Kb 6.2 Kb A B C Spleen Thymus Prostate Testis Ovary Small Intestine Colon Peripheral blood Leukocyte Heart Brain Placenta Lung Liver Skeletal Muscle Kidney pancreas FXAML-LJ: IDENTIFICATION OF HUMAN OB RECEPTOR GENOMIC DNA AND CHROMOSOME LOCALIZATION; IDENTIFICATION OF HUMAN OB RECEPTOR "D"n The full length human OB receptor genomic DNA was also prepared. OB receptor "A"l cDNA, in its entirety, was used as a probe against a human genomic DNA library, using materials and methods from a commercially available kit (Genome Systems, using a human genomic library in a P1 vector) A single 47 positive clone was detected. There are introns located at (with respect to OB receptor DNA) base pair number: 559, 1059, 1350, 1667, 1817, 1937, 2060, 2277, 2460, 2662, and 2738.

The human OB receptor gene was localized to human chromosome 1P31 by FISH analysis (Genome Systems).

Human chromosome 1 is thought to correspond to mouse chromosome 4C7, which is presumed to be the location of the db locus.

A further chromosomal sequence was isolated.

This chromosomal DNA sequence was isolated from a human genomic library as described above. This chromosomal sequence encodes what is here denominated human OB receptor and the encoded amino acid sequence is set forth in SEQ. ID No. 7. A cDNA encoding this amino acid sequence is set forth in SEQ. ID No. 8. The chromosomal DNA intron/exon junction map is set forth as SEQ. ID No. 9.

As with forms and for the S" 20 present form OB receptor protein, the first amino 00 acid of the mature protein is likely (using Shydrophobicity analysis) to begin at position 22 23 or 29 The last amino acid of the protein is at position 1165 and is a valine residue. As with the other forms, the extracellular domain extends from position 22 23 or 29 to position 839 or *0o. 841 The transmembrane domain appears to begin at position 840 or 842 The end of the transmembrane domain appears to be located at position 30 862 863 or 864 The C-terminal region, beyond the transmembrane region, is likely to be involved in signal transduction, and is located at position 863 864 or 865 through position 1165 The present OB receptor form is identical to that published by Tartaglia et al, Cell 13: 1263-1271 48 (December 29, 1995) with the exception of a single amino acid change at amino acid position 976 (nucleotide codon begining at position 3022). The present type amino acid at position 976 is aspartic acid, and the published amino acid corresponding to the same position is alanine. This is a non-conservative substitution, see infra, and since the location of the substitution is within a region thought important for signal transduction, this change could affect the function of the molecule.

EXAMPLE 4: PREPARATION OF SOLUBLE OB RECEPTOR Three forms of soluble human OB receptor have been prepared: 1. Leader Extracellular Domain (Seq.

ID Nos. 10 and 11): A recombinant form of the soluble human OB receptor was prepared. This form encompasses, in the immature protein, the leader sequence and the 20 extracellular domain (amino acids 1-839). The mature protein would have the leader sequence deleted, and the first amino acid of the mature recombinant soluble human OB receptor would be 22 23 or 29 This protein was expressed as described below.

25 2. Leader Extracellular Domain Cterminal FLAG (Seq. ID No. 12): A second form of the "recombinant soluble human OB receptor was also prepared.

This form had a "FLAG" tag located at the terminus of the protein. The "FLAG" peptide is a useful research tool as it allows one to follow the protein using an antibody which recognizes the "FLAG" peptide. Such reagents are commercially available (IBI, New Haven, CT). This protein was expressed as described below.

3. Native Splice Variant (Seq. ID Nos.

13 and 14): This form is believed to the the recombinant form of a naturally occurring secreted, 49 soluble human OB receptor. This form has most of the amino acids found in the extracellular domain (amino acids 22-798), and a unique 6 amino acid sequence at the carboxyl terminus. Beginning at amino acid position 799 of Seq. ID No. 13, the amino acid sequence of this native splice variant human OB receptor protein is "G K F T I L." EXAMPLE 5: PREPARATION OF EXPRESSION VECTORS Recombinant human OB receptor expression vectors have been prepared for expression in mammalian cells. As indicated above, expression may also be in non-mammalian cells, such as bacterial cells. The type cDNA (Seq. ID No. 2) was placed into a commercially available mammalian vector (pCEP4, Invitrogen) for expression in mammalian cells, including the commercially available human embryonic kidney cell line, "293".

Recombinant human OB receptor expression 20 vectors have been prepared for expression of recombinant soluble OB receptor, consisting of the leader sequence and the extracellular domain (Seq. ID Nos. 10 and 11), using the same system as above (the commercialy available mammalian vector pCEP4, and "293" cells).

This recombinant soluble human OB receptor was.also expressed in CHO cells in a similar way.

The "FLAG-tagged" form (Seq. ID No. 12) of the recombinant soluble human OB receptor, ahd the form (Seq. ID No. 7) were also expressed in "293" cells in a 30 similar fashion as above.

Detection of desired protein was accomplished using BIACORE (Pharmacia) analysis. This analysis is analogous to that described in Bartley et al., Nature .36: 558-560 (1994).

Essentially, the BIACORE machine measures affinity interactions between two proteins. In this 50 case, the OB protein was immobilized on the machine, and conditioned media from cell lines expressing the OB receptor was added to the machine. Any receptor protein present in the conditioned media bound to the OB protein surface. The BIACORE machine gave a read-out indicating that receptor protein was being expressed. For recombinant soluble receptor (Seq. ID No. 10) expression in "293" cells, the read-out was 191.0 relative to a baseline readout of 0. For recombinant soluble receptor (SEq. ID No. 10) expression in CHO cells, the read-out was 150.9 relative to a baseline readout of 0. For recombinant soluble receptor with a C-terminal FLAG-tag (Seq. ID. No. 12), the read-out was 172.0 relative to a baseline of 0.

For expression in bacterial cells, one would typically eliminate that portion encoding the leader sequence potentially amino acids 1-21, 1-22 or 1- 28). One may add an additional methionyl at the N-terminus for bacterial expression. Additionally, one may substitute the native leader sequence with a different leader sequence, or other sequence for •cleavage for ease of expression.

EXAMPLE 6: DEMONSTRATION OF SIGNAL TRANSDUCTION This example demonstrates that the form is active to produce a signal within a cell, whereas in the same cell type, the form does not. The signal transduction assay was performed by the use of "293" cells transiently expressing either the or the "D" form (see above for preparation of the "293" expression clones) Phosphorylation of molecules predicted to be involved in signal transduction within the cell was examined upon OB protein binding to the OB receptor protein tested. The results demonstrate that upon binding of OB protein to the extracellular domain, the 51 form of the present OB protein receptor transduces a signal sufficient to initiate phosphorylation of signalling molecules.

Methods 1. OB receptor molecules. As indicated above, the form (Seq. ID No. 1) and the form (Seq. ID. No. 7) were studied.

2. Expression system. The pCEP 4 system (as described above) having inserted DNA encoding the "A" form (Seq. ID No. 2) or the form (Seq. ID No. 8) was used to transfect "293" cells. These cells did not allow for the pCEP4 vector to integrate into the genome, so such expression was transient. Non-recombinant (mock-transfected) cells were also prepared as controls.

3. Detection of phosphorylation. Mock transfected cells and cells expressing the form or the form were analyzed. Prior to treatment the cells were serum-starved by incubation in media with 0.5% serum for 16 hours prior to the treatments. The cells were treated with the OB protein (10 mg/ml) for minutes at 37 0 c, after which the cells were lysed in modified NP40 buffer (50 mM Tris, pH 8.0, 150 mM sodium chloride, 1% NP40, 10 mg/ml aprotinin, 5mM EDTA, 200 mM sodium orthovanadate). Phosphotyrosine containing proteins were immunoprecipitated (Anti-phosphotyrosine antibody 4G10, UBI, Lake Placid, NY), and separated by SDS polyacrylamide gel electrophoresis. After electrophoresis and electroblotting to membranes the 30 immunoprecipitates were probed with antibodies to various signal transduction.molecules. Antibodies to STATs, JAKs and ERKs were purchased from Santa Cruz Biotechnology Inc. Immune complexes were detected by horseradish peroxidase conjugated secondary reagents using chemiluminescence as described by the manufacturer (ECL, Amersham). As a positive control, 32D cells were 52 treated with IL-3, which is known to activate by tyrosine phosphorylation most of the molecules being analyzed.

4. Results. Results are presented in Table 3, below. As can be seen, only the form was able to respond to either mouse or human OB protein as detected by phosphorylation of JAK and STAT molecules. A designation indicates signal was detected, a designation means that no signal was observed.

TABLE 3 Signal 293 293/D 293/D 293/A 293/A 32D /ABt Alone hrOB* mrOB** hrOB# mrOB## IL-3 STAT1 STAT3 STATS JAK1 JAK2 JAK3 TYK2 ERKs 1,2 t Antibody detection target 293 cells expressing receptor form treated wit 15 recombinant human OB 293 cells expressing receptor form treated with recombinant murine OB 293 cells expressing receptor form treated with recombinant human OB 293 cells expressing receptor form treated with recombinant murine OB The form is capable of initiating signalling through the JAK/STAT pathways in 293 cells, whereas the form cannot.

h 1 1 l 53 EXAMPLE 7: USE OF SOLUBLE OB RECEPTOR AS A THERAPEUTIC This example demonstrates that soluble OB receptor protein acts to protect the activity of OB protein. Below, soluble OB receptor and/or OB protein was delivered to a mammal via "gene transplant" that is, via bone marrow cells engineered to express the desired DNAs. When soluble OB receptor combined with OB protein was delivered, the animals lost more weight than delivery of OB protein alone. This demonstrates the protective activity of OB receptor protein.

While not wishing to be bound by theory, one explanation of the mode of action is that soluble OB receptor protein acts to protect the OB protein in serum from agents or conditions which could diminish its activity. The protective action appears to increase circulating half-life of the protein. As such, the present example demonstrates that OB receptor either 20 alone, or administered as a complex with OB protein (or analog or derivative thereof) could act as a therapeutic agent.

Materials and methods: 1. Preparation of recombinant ob retroviral vector Packaging Cells.

.Use of murine ob cDNA. Full length wild-type murine ob cDNA was amplified by the PCR using synthetic oligonucleotides designed from the published sequence Zhang et al., Nature 372. 425-432 (1994) .Linkers (An Eco RI linker and a B _l II linker) were used to facilitate subcloning.

Use of soluble recombinant human OB receptor CDNA. Methods similar to those above were used. A construct containing the recombinant human soluble receptor of Seq. ID No. 10 was used, and modified with 54 linkers to facilitate cloning the addition of a Bg1 II restriction endonuclease recognition site).

Placement of desired cDNA into vector. PCR products were digested with EcoRI and BglII and cloned into similarly-digested parental vector (pMSCV2.1) under the transcriptional control of the viral LTR promoter.

The parental MSCV vector (supplied by R. Hawley, University of Toronto, Canada) was derived from MESV (murine embryonic stem cell virus) and contains a neomycin phosphotransferase resistance (neor) gene driven by an internal mouse phosphoglycerate kinase (PGK) promoter, as described- Hawley, et al, J. Exp.

Med. 176: 1149 -1163 (1992). The parental plasmid pMSCV2.1 and pMSCV-OB were independently electroporated into the GP+E-86 packaging cell line (supplied by Dr. A.

Bank, Columbia University, NY) Markowitz et al., J.

Virol. 62:1120-1124 (1988). Transient supernatants were harvested from electroporated populations and used to infect tunicamycin treated parental GP+E-86 cells.

20 Tunicamycin treatment relieves the block to superinfection of the parental packaging cells. G418 S•(0.78 mg/mL, 67% active, GIBCO Laboratories, Life Technologies, Inc., Grand Island, NY) resistant clones were selected from each infected population and titered by infection of NIH3T3 cells. Clones with the highest G418 resistant titer were expanded and frozen as S" aliquots. Each bone marrow infection and transplantation experiment used aliquots from the same passage of frozen viral packaging cells. Both the parental and ob packaging cell lines were tested for the presence of, and found to be free from, replication competent virus using a sensitive marker rescue assay.

Moore, et al., (1993) in: Gene Targeting: A Practical Approach, Joyner, Ed. (Oxford University Press, New York, NY).

55 2. Production of Retroviral SuDernatants, Recombinant virus-producing packaging cell lines were grown in 175cm 2 tissue culture flasks in Iscove's Modified Dulbecco's Medium (IMDM) (GIBCO), 10% (v/v) FBS, at 37 0 C. Sub-confluent (approximately monolayers of cells were fed with fresh medium 24h prior to harvest of virus-containing supernatants. Viral supernatants were removed from packaging cell lines by aspiration, sterile filtered (0.45mM) and added directly to bone marrow cultures. Fresh aliquots of frozen packaging cell lines were thawed for use in each experiment.

3. Bone Marrow Infection and Transplantation.

Eight to 12-week old female C57BL/6J or (ob/ob) mice were used as bone marrow donors and recipients.

All mice were purchased from The Jackson Laboratory (Bar Harbor, ME) and housed under specific pathogen-free conditions in a vivarium in accordance with governmental regulations and institutional guidelines.

20 Bone marrow cells were harvested from femurs S.and tibias of donor mice 4 days post 5-fluorouracil FU, Sigma Chemical Co., St. Louis, MO) treatment (150 mg/kg Bone marrow cells (6 X 10 5 /mL) were incubated in 150mm tissue culture dishes containing fresh viral supernatant (as described above), 15% FBS, 6 mg/mL polybrene (Sigma), 0.1% bovine serum albumin (BSA, Fraction V, Sigma), 2.5 ng/mL recombinant mouse IL-3 (rmIL-3), 100 ng/mL each of recombinant human IL-6 (rhIL-6), recombinant human IL-11 (rhIL-1l), and 30 recombinant rat SCF (rrSCF). All growth factors were produced by Amgen, Inc. (Thousand Oaks, CA). Culture media were replaced daily for 3 days with fresh viruscontaining supernatant and growth factors.

At the end of the infection period, total nonadherent and adherent cells were washed and resuspended in 1% BSA-saline and transplanted into g-irradiated (12 56 Gy, Cs 137 mice. Each animal was transplanted with X 10 6 syngeneic cells. There were approximately animals per cohort.

4. Analysis of OB protein expression in transfected cells and transplanted animals. For transfected bone marrow cells, Western analysis was performed. Vector packaging cell supernatant was resolved by SDS-PAGE (16% acrylamide), then transferred to Hybond-ECL (Amersham, Arlington Heights, IL). The filter was incubated with affinity-purified rabbit amouse OB protein polyclonal antibody (1mg/mL) in T-TBS buffer (20mM Tris-chloride, pH7.6, 137mM NaC1, 0.1% at room temperature for 45 min. Horseradish peroxidase (HRP)-conjugated donkey a-rabbit IgG (Amersham) was diluted in T-TBS (1:2500) and incubated with the filter at room temperature for 45 min.

Enhanced chemiluminescence (ECL, Amersham) detection was performed as recommended by the manufacturer.

For transplanted animals, serum was analyzed.

20 Animals were bled retroorbitally, under isofluorane anesthesia. Serum from transplanted ob/ob animals was resolved by SDS-PAGE (4-20% acrylamide) under nonreducing and reducing conditions, then transferred to Trans-Blot (Bio-Rad Laboratories, Hercules, CA) membranes. The membranes were incubated for 2 hours at room temperature with HRP-conjugated rabbit a-mouse OB protein antibody (0.125mg/mL) in T-TBS buffer containing fetal bovine serum and 1% bovine serum albumin.

Bound OB protein was detected by ECL (Amersham), 30 performed as recommended by the manufacturer.

For quantitation of soluble OB protein levels, serum from transplanted animals was subjected to ELISA analysis. Briefly, affinity-purified rabbit a-OB protein polyclonal antibody was coated onto 96-well plates. Standards (purified recombinant OB protein 57 monomer, Pelleymounter et al., Science 269: 540-543 (1995) and experimental samples were added, and the plates were incubated at room temperature. The plates were washed twice and affinity-purified rabbit a-OB protein antibody conjugated to horseradish peroxidase was added. Following incubation at room temperature, the plates were washed four times with TMB/peroxide substrate was added and the color reaction was read at 450nm in a Molecular Devices plate reader.

OB protein concentrations in sera were estimated by comparison to a standard curve prepared from internal standards. OB protein levels were reliably measured in samples containing >160 pg/mL.

Body Weight and Food Intake. Mice were offered pelletized rodent chow (PMI Feeds, Inc., St.

Louis, MO) ad libitum. The body weight of individual animals was measured daily for the first two months of 00. analysis, and weekly thereafter. Food consumption was measured daily on selected groups of individually-housed 20 animals.

0 Results Results are presented in Tables 4 and 5 below.

Administration of OB protein receptor increased the effectiveness of OB protein. This may have been accomplished via an increased circulation time of OB .00. protein in the presence of OB protein receptor.

As can be seen in the Table, animals administered a combination of OB protein and OB protein 0'0 30 receptor (via genetic therapy) had a greater weight loss after 28 days than either composition alone. The Table presents the results of two experiments As can be seen, use of the OB protein alone at day resulted in animals with 87.5% and 72.2% of the starting weight. Using OB receptor in combination with OB protein, however, resulted in animals with 68% and 58 53.6% of the starting weight. Use of the receptor alone appeared to have little effect, if any.

TABLE 4 Treatment Weight(g) starting starting decrease at weight weight day 28 (ave) (ave) (ave) day 28 day OB alone* 6.3/12.7 87.9/75.3 87.5/72.2 Receptor** 103/100.6 104.2/101.7 alone OB 12.6/16.8 76.3/67.5 68/53.6 Receptor***, 50% bone marrow cells transfected with OB protein cDNA as described above, and 50% bone marrow cells without genetic alteration 50% bone marrow cells transfected with OB receptor 10 protein cDNA as described above, and 50% bone marrow cells without genetic alteration 50% bone marrow cells transfected with OB protein cDNA as described above, and 50% bone marrow cells transfected with OB receptor protein cDNA as described above.

e* Table 5, below, contains results of the OB levels found in the serum from animals administered OB protein alone, or administered OB protein in combination with OB protein receptor (via the "gene therapy" method 20 of this example). The data reflect nanograms of OB protein per milliliter of serum, plus or minus the standard error of the mean.

*00* 59 TABLE Treatment Experiment #1 Experiment #2 t OB alone* 2.93 0.77 9.74 1.02 Receptor** 0.08 0.05 0.12 0.07 alone OB 12.11 1.90 15.18 2.52 Receptor*** 50% bone marrow cells transfected with OB protein cDNA as described above, and 50% bone marrow cells without genetic alteration 50% bone marrow cells transfected with OB receptor protein cDNA as described above, and 50% bone marrow cells without genetic alteration 50%.bone marrow cells transfected with OB protein cDNA as described above, and 50% bone marrow cells transfected with OB receptor protein cDNA as described above.

Experiment #1 was conducted as described above, 15 with OB protein serum levels measured after 38 days.

t: Experiment #2 was also conducted as described above, with OB protein serum levels measured after 24 days.

The data demonstrate the protective effects of OB receptor. As can be seen, in the presence of OB receptor, OB protein has a higher accumulation in the serum. The degree of accumulation is observed to increase inversely with the levels of OB protein in the serum. In Experiment #1 (with a base OB protein level of about 2.93 ng/ml), the OB protein serum level increased about 400% with the addition of receptor, where in Experiment #2 (with a base of about 9.74), the OB protein serum level increased by about OB receptor administered either alone or in association with OB protein (or analogs or derivatives 60 thereof) may serve to increase the circulation time of OB protein, and therefore enhance the therapeutic efficacy of either exogenous or endogenous OB protein.

EXAMPLE 8: PREPARATION OF SELECTIVE BINDING MOLECULES Animals were immunized for the preparation of polyclonal antibodies using the following peptides (with respect to the numbering of the amino acids for OB receptor A, Seq. ID No. 54-64; 91-100; 310-325; 397-406; 482-496; 874-885; and, with respect to amino acids of OB receptor (Seq. ID No. 910-929. Some of the polyclonal antibodies prepared (in rabbits) were tested for ability to bind to recombinant human OB receptor protein. The polyclonal antibody prepared against amino acids 54-64 was found to have the highest affinity for recombinant human OB receptor protein. The polyclonal antibody prepared against amino acids 397-406 was also found to bind to recombinant human OB receptor protein. The polyclonal antibody prepared against amino 20 acids 91-100 was found to slightly bind to recombinant human OB receptor protein. The polyclonal antibody *i prepared against amino acids 874-885 was found not to bind to recombinant human OB receptor protein.

An additional study was performed which 25 demonstrates the expression and purification of the extracellular domain of the OB receptor protein in CHO cells, and antibodies which recognize this OB protein receptor extracellular domain.

The extracellular domain of the human OB receptor protein was expressed as a secreted, soluble protein in CHO cells as previously described supra.

Individual cell lines were isolated and grown in increasing amounts of methotrexate to increase selection/expression of the recombinant receptor protein (100, 200 or 500 micrograms methotrexate per ml of media). Conditioned media from the CHO cell lines was 61 collected, and the proteins in the conditioned media were fractionated by SDS-PAGE. The OB receptor extracellular domain migrated as a broad band with an apparent size range of about 140 kDa to about 200 kDa.

The OB receptor protein extracellular domain was detected by Western Blot analysis using polyclonal antibodies prepared against a portion of the extracellular domain of the OB receptor protein. The unfolded, bacterially expressed protein was used as an antigen to generate antisera in rabbits. The identified OB receptor extracellular domain was purified by affinity chromatography. The purified protein was sequenced at the amino terminus to confirm that it was the OB receptor and also to determine the start of the mature protein (after signal peptide cleavage) as expresed in CHO cells. It was found that amino acid no. 22 (according to the amino acid sequence numbering of Seq.

ID No. 1, infra), was the first amino acid of the mature protein as expressed in CHO cells.

20 Other immunogenic peptides may be used.

Polyclonal, monospecific polyclonal, monoclonal, antibody fragments, and recombinant antibodies may be prepared using methods available to those skilled in the art.

25 One may further use recombinant techniques or peptide synthesis methods to alter the character of such selective binding molecules. This may be accomplished by preparing recombinant antibodies having altered complementarity determining regions (sometimes referred o 30 to in the art as "CDR's") to, for example "humanize" the antibodies by using human Fc (constant) regions. Other types of recombinant antibodies, for example, those having CDR's altered to enhance affinity or selectivity to one or more members of the OB receptor family, may be prepared and used using methods available to those 62 skilled in the art. See Winter et al., Nature 349: 293- 299 (1991).

The present OB receptor protein may be used as an assay to screen for desired selective binding molecules. Such assay may be based on binding capability, or biological activity, or, other means of detecting signal transduction. For example, if one were to prepare a series of modified antibodies, one could test them for affinity binding strength) against the target OB receptor.

The selective binding molecules may be useful for diagnostic purposes, such as tissue distribution analysis, or to diagnose the relative affinity of an individual's OB receptors for such selective binding molecule to determine the functionality of an individual's OB receptor during a course of therapy.

Selective binding molecules may be alternative therapeutic or cosmetic products to OB protein.

20 EXAMPLE 9: GENE THERAPY One may deliver the present OB receptor .protein via gene therapy, as described infra.

One may envision, using materials and methods available to those skilled in the art and provided herein, using T-cells as an agent carrying DNA expressing OB receptor for gene therapy. An individual would have T-cells selected using CD34+ selection and a magnetic microparticles selection device. Such cells would be transfected with the desired DNA, or the regulation of the desired coding region may be altered using homologous recombination or other in situ techniques. The transduced cells could be selected empirically, using means to detect the desired protein, or a marker may be included which permits indirect detection a selectable marker as is known in the 63 art). Optionally, such cells could be expanded, for example, using one or more growth factors such as SCF or an interleukin, and such cells could be stored for future use. In such a way, the procedure would only have to be accomplished once or infrequently in an individual's lifetime, for later transfer into the individual. The cells would be re-planted into the individual, and the individual would be monitored for desired therapeutic effect, such as weight loss/maintenance of weight, diabetes recurrence, blood lipid levels, or other conditions.

Illustrative Nucleic Acid and Amino Acid Sequences The below amino acid and DNA sequences are those to which reference has been made. An asterick("*") indicates the position of a stop codon.

64 Human OB Rece-otor Amino Acid Seqruence (Seg. -ID No. 1 (Amino Acid. single letter abbreviation)o 1 51 101 L 0 151 201 251 L 301 351 401 451 501 551 601 30 651 701 751 801 851 901 951

MICQKFCVVL

AGL SKNT SN S LCADN IEGKT

LFRNYNYKVH

P TAKLNDTLL GNLKI SWSSP GS SYEVQVRG

HCIYKKENKI

PRGKFTYDAV

TSTIQSLAES

QP IFLLSGYT

IGLLKISWEK

P DLCAVYAVQ

GDTMKKEKNV

FTFLWTEQAH

SSCVIVSWIL

FIPIEKYQFS

SSSILLLGTL

MITTDEPNVP

T*NRCVNLGS

LHWEFIYVIT AFNLSYPITP WRFKLSCMPP NGHYETAVEP KFNSSGTHFS NLSKTTFHCC FVSTVNSLVF QQIDANWNIQ CWLKGDLKLF LLYVLPEVLE DSPLVPQKGS FQMVHCNCSV MCLKITSGGV IFQSPLMSVQ PINMVKPDPP PLVPFPLQYQ VKYSENSTTV IREADKIVSA KRLDGPGIWS DWSTPRVFTT QDVIYFPPKI VPSKEIVWWM NLAEKIPQSQ YDVVSDHVSK YCCNEHECHH RYAELYVIDV NINISCETDG TLQLRYHRSS LYCSDIPSIH PISEPKDCYL MWIRINHSLG SLDSPPTCVL PDSVVKPLPP PVFPENNLQF QIRYGLSGKE VQWKMYEVYD VRCKRLDGLG YWSNWSNPAY TVVMDIKVPM TLLWKPLMKN DSLCSVQRYV INHHTSCNGT TVTVLAINSI GASVANFNLT FSWPMSKVNI SPSDYKLMYF IIEWKNLNED GEIKWLRISS LYPIFMECVG KPKIINSFTQ DDIEKHQSDA LISHQRMKKL FWEDVPNPKN CSWAQGLNFQ TSQQSIEY*K IFTF*RRGAN ILKKIQLNF*E KCPJESSLDV *L

NSTYDYFLLP

FRSEQDRNCS

ICYVESLFKN

HECCECLVPV

LGLHMEITDD

TSLLVDSILP

LTSVGSNVSF

VTFFNLNETK

YLTKMTCRWS

QSDGFYECIF

S SVKAE ITIN

AKSKSVSLPV

RGPEFWRI IN

WSEDVGNHTK

VQSLSAYPLN

SVKKYYIHDH

GLYVIVPVI I

KRTDIL*SLI

LTYGGLC *FR 65 Human OB Recelptor DNA Secauence (Secr. !D No. 2 (DNA) 000.

0 0 0 1 51 101 151 LO0 201 251 301 351 401 451 501 551 601 651 701 751 801 851 901 951 1001 1051 1101 1151 1201 1251

CCGCCGCCAT

CTTC TCTGAA

TTGGGAATTT

CTCC TTGGAG TACT TCCT TT ACAT TAT GAG

TTTCTAACTT

GATAGAAACT

TTCAACAGtA

TACAGTGCTG

TCATTATTTA

ATATGTTCTG

GCAGTTTTCA

TGTCTTGTGC

TTTGAAAATC

TTCAGCCC AT

GAAATCACAG

GGTACCATTT

CAGT TATCAG

GACAGTATAC

ACTGGATGGC

CCACACAAGA

TCTAATGTTT

CTCAAAAGAG

GCCAGTATGA

CTGAATGAAA

CTCTGCCTTC

GTAAGATGAT

ATTTATGTGA

ATTTAAGTTG

TGCCTGCTGG

ACAGCTGTTG

ATCCAAAACA

GCTCCTTATG

AATTCTTTAG

GCTAAAAGGA

AGAATCTATT

CCTGAAGTGT

GATGGTTCAC

CTGTGCCAAC

ACATCTGGTG

AAATATGGTG

ATGATGGTAA

CCACTTCAAT

AGAAGC TGAC

TTCCTGGGTC

CCAGGAATCT

TGTCATATAC

CTTTTCACTG

ATTGTTTGGT

TGTTGTGAGT

CCAAACCTCG

GGTCGAG TTG

TTGTCAAAAA

TAACTGC GTT

TCTTGCATGC

ACTCTCAAAG

AACCTAAGTT

ACTTTCCACT

T GCAGACAAC T TTTTCAACA

GACTTAAAAT

CAGGAATTAT

TAGAAGATTC

TGCAATTGCA

AGCCAAACTC

GAGTAAT TTT

A.AGCCTGATC

TTTAAAGATT

ATCAAGTGAA

AAGATTGTCT

T TCGTATGAG

GGAGTGACTG

TTTCCACCTA

CATCTATAAG

GGATGAATTT

GATCATGTTA

AGGAAAGTTT

GACCCCCGGA

TTCTGTGTGG

TAACTTGTCA

CACCAAATTC

AATACTTCA.A

TAATTCAAGT

GTTGCTTTCG

ATT GAAGGAA

AATAGATGCA

TATTCATCTG

AACTATAAGG

ACCTCTGGTT

GTGTTCATGA

AACGACACTC

CCAGTCACCT

CACCATTAGG

TCTTGGTCCA

ATATTCAGAG

CAGCTACATC

GTTCAGGTGA

GAGTACTCCT

AAATTCTGAC

AAGGAAAACA

AGCTGAGAAA

GCAAAG.TTAC

ACCTATGATG

TCAAGGTGTA

TTTTGTTACA

TATCCAATTA

AACCTATGAC

ATTCGAATGG

GGTACTCACT

GAGTGAGCAA

AGACATTTGT

AAC TGGAACA

TTATGTGGAG

TCCATCTTTT

CCCCAAAAAG

ATGTTGTGAA

TCCT TATGTG

CTAATGTCAG

TTTGCATATG

GCCCACCATT

AATTCTACAA

CCTGCTAGTA

GGGGCAAGAG

CGTGTCTTTA

AAGTGTTGGG

AGATTGTTCC

ATTCCTCAAA

TTTTTTCAAT

CAGTGTACTG

66 1301 1351 1401 1451 1501 1551 1601 1651 1701 1751 1801 1851 1901 1951 30 2001 2051 2101 2151 2201 40 2251 2301 2351 2401 2451 2501

CTGOAATGAA

ATGTCAATAT

ACTTGCAGAT

GCAATTGAGG

TTOATCCCAT

TATGAATGOA

GAT TAO GATO TOOTTOO TOA

OAAATTAOTA

OTTTOOAGAG

AAGAAGTAOA

GTOAGTOTOO

OTGTAAGAGG

OOTAOAOAGT

TGGAGAATAA

AOTTTGGAAG

ATGTGATAAA

GGAAATOAOA

TAOGGTTOTG

TAACCTTTTO

GOTTATOOTT

CAGT GAT TAO

AAGATGGTGA

TATATCOATG

CCCAATATTT

OATGAATGOO

OAATATOTOA

GGTOAAOOAG

TATOATAGGA

ATOTGAGOOO

TTTTOOAGOO

AATOAOTOTO

TTOTGTGGTG

TAAACATTG

AATAAOOTTO

ATGGAAGAT G

OAGTTOOAGA

O TAGAT OOAO

TGTOATGGAT

TTAATOOAGA

OOOOTGATGA

OOATOATACT

OGAAATTOAO

GOOATOAATT

AT G 00 TAT G

TAAAOAGCAG

AAGOTAATGT

AATAAAATGG

ATOATTTTAT

ATGGAAGGAG

ATOATOGOTA

TOTGAAAOTO

TAOAATOOAG

GOAGOOTTTA

AAAGATTGOT

AATOTTOOTA

TAGGT TOAOT

AAOOOAOTGO

ATTATTGAAA

AATTOOAGAT

TATGAGGTTT

OTTGTGTGOA

TGGGATATTO

ATAAAAGTTO

TAOTATGAAA

AAAATGAOTO

TOOTGOAATG

TTTCOTGTGG

OAATTGOTGO

AGOAAAGTAA

TTGTGTGATT

ATTTTATTAT

CTTAGAATCT

CCOATTGAG

TGGGAAAACC

TOO TOAATTA

ATGOGTAOTT

TOAOTTGOGG

OTGTTOTGAT

ATTTGOAGAG

TTATOTOGOT

TOACTOTOOA

OTOOATOOAG

ATATOTTGOG

TOGOTATOOT

ATOATGOAAA

GTOTATGOTG

GA TA.ATTGO

OTATGAGAGG

AAGOAGAAAA

ATTGTGOAGT

GAAOATGGTO

AOAGAGOAAG

TTCTGTTGOA

ATATOGTGOA

GTTTOOTGGA

TGAGTGGAAA

OTT OATOTGT

AAGTAOOAGT

AAAGATAAT T

TATGTOATTG

AAOTAAAATO

AAAOOAOTTT

ATTOOATCTA

TGAT GGT T TT

AOAOAATGTG

OOAAOATGTG

TOTGAAAGOA

AAAAGOOAGT

TTAAGTGGAA

ATOAAAATOT

TTOAGGTGOO

AGOAATOOAO

AOOTGAATTT

ATGTOAOTTT

OT TOAGAGAT

AGAAGATGTO

OAOATAOTGT

AATTTTAATT

GTOAOTOAGT

TACTATCACO

AATOTTAATG

TAAGAAG TAT

TOAGTOTTTA

AATAGTTTCA

2551 OTOAAGATGA TATTGAAAAA OAOOAGAGTG ATGOAGGTTT ATATGTAATT 67 2601 2651 2701 2751 2801 LO0 2851 2901 L 5 2951 3001 3051 3101 3151

GTGCCAGTAA

ATCACACCAA

AGAATTGTTC

CTTTGAAGTC

CCAACAGTCT

CAAATCTAAA

TTATGTTGAT

TAGATTTGAG

TTTAAAAGTA

TCATAAAACA

AAATGTCATC

AACTGCAACA

TTATTTCCTC TTCCATCTTA TTGCTTGCAA CATTATTAAT AGAATGAAAA AGCTATTTTG GGAAGATGTT CCGAACCCCA CTGGGCACAA GGACTTAATT TTCAGAAGAG AACGGACATT TAATCATGAT CACTACAGAT GAACCCAATG TGCCAACTTC ATAGAGTATT AGAAGATTTT TACATTTTGA AGAAGGGGAG AAAAATTCAG TTGAACTTCT GAGAGTTAAC ATATGGTGGA TTAGAACTTA AAATAGATGT GTAAATTTGG GTTCAAAATG TCCAGTTTGG ATGTGTGATT AATTTTCAA.A TCATCTAA.AG GTATTCATGA TTTCTGGCTT TTGATTTGCC ATATTCCTGG TTAAGAAAAT TATGGCTGTT GCTGTCATTA CATATCTATT AAATATGTAG TAGACAATTT TGTAATTAGG TGAACTCTAA TCTGACAAAT TGCTTTAAAA ATACAATGAT TAT 9* 68 Human OB Receotor Amino Acid Seauipnc'p (.qscT Tfl Nco thi 1 51 101 151 201 251 301 351 401 451 25 501 551 601 651 701 35 751 801 851 901 951 MI CQKFCVVL

AGLSKNTSNS

LCADN IEGKT

LFRNYNYKVH

PTAKLNDTLL

GNLKI SWS SP

GSSYEVQVRG

HCIYKKENKI

PRGKFTYDAV

TSTIQSLAES

QPIFLLSGYT

IGLLKISWEK

PDLCAVYAVQ

GDTMKKEKNV

FTFLWTEQAfH

SSCVIVSWIL

FIPIEKYQFS

SSSILLLGTL

IQHQ*HVVLF

LFVLVTS STV LHWEFIYVIT AFNLSYPITP NGHYETAVEP KFNSSGTHFS FVSTVNSLVF QQIDANWN IQ LLYVLPEVLE DSPLVQKGS MCLKITSGGV IFQSPLMSVQ PLVPFPLQYQ VKYSENSTTV KRLDGPCIWS DWSTPRVFTT VPSKEIVWWM NLAEKIPQSQ YCCNEHECHH RYAELYVIDV TLQLRYHRSS LYCSDIPSIH- MWIRINHSLG SLDSPPTCVL PVFPENNLQF QIRYGLSGKE VRCKRLDGLG YWSNWSNPAY TLLWKPLMKlN DS LCSVQRYV TVTVLAINS I GASVANFNLT SPSDYKLMYF I IEWKNLNED LYPIFMEGVG KPKIINSFTQ LISHQRMKKL FWEDVPNPKN FWSLKQFQKI SVLIHHGKIK LT SLRLRVLR PMRTKARDN WRFKLSC2PP

NLSKTTFHCC

CWLKGDLKLF

FQMVHCNCSV

P INNVKPDP I READK IVSA QDVI YFPPKI YDVVS DHVS K

NINISCETDG

PT SEPKDCYL

PDSVVKPLPP

VQWKMYEVYD

TVVMD IKVPM

INHHTSCNGT

FSWPMSKVNI

GE IKWLRIS S DD IEKHQSDA

CSWAQGLNFQ

M*CQQLWSL

PLLNTPR* SA

NSTYDYFLLP

FRSEQDRNCS

ICYVESLFKN

HECCECLVPV

LGLHMEITDD

TSLLVDSILP

LTSVG SNVSF VT FEN LNET K

YLTKMTCRWS

QSDGFYECIF

S SVKAE ITT N

AKSKSVSLPV

RGPEFWRI IN

WSEDVGNHTK

VQSLSAYPLN

SVKKYY IHDH GLYVI VP VTI KKRLS IFLSS YFQQQ ILKRV

TLNQVKLVK

@O9S 0*O* 0 @900 0* 99 0 0 0 *9 9 5 0 0 *5*0 9 *9 0 9* 5* 5*@0 69 Human OB Recep2tor DNA Seauence (Sea., ID No. 4 (DNA)): 1 52.

1021 151 201 251 301 351 401 451 501 551 601 651 701 751 801 851 901 951 1001 1051 1101 1151 1201 1251 COGCGGCAT CTCTGCCTTC CTTCTCTGAA GTAAGATGAT TTGGGAATTT ATTTATGTGA CTCCTTGGAG ATTTAAGTTG TACTTCCTTT TGCCTGCTGG ACATTATGAG ACAGCTGTTG TTTCTAACTT ATCCAAAACA GATAGAAACT GCTCCTTATG TTCAACAGTA AATTCTTTAG TACAGTGCTG GCTAAAAGGA TCATTATTTA AGAATCTATT ATATGTTCTG CCTGAAGTCT GCAGTTTTCA GATGGTTCAC TGTCTTGTGC CTGTGCCAAC TTTGAAAATC ACATCTGGTG TTCAGCCCAT AAATATGGTG GAAATCACAG ATGATGGTAA GGTACCATTT CCACTTCAAT CAGTTATCAG AGAAGCTGAC GACAGTATAC TTCCTGGGTC ACTGGATGGC CCAGGAATCT CCACACAAGA TGTCATATAC TCTAATGTTT CTTTTCACTG CTCAAAAGAG ATTGTTTGGT GCCAGTATGA TGTTGTGAGT CTGAATGAAA CCAAACCTCG GGTCGAG TTG

TTGTCAAAAA

TAACTGCGTT

TCTTGCATGC CACCAAATTC ACTCTCAAAG AATACTTCAA AACCTAAGTT TAATTCAAGT ACTTTCCACT GTTGCTTTCG TGCAGACAAC ATTGAAGGAA TTTTTCAACA AATAGATGCA GACTTAAAAT TATTCATCTO CAGGAAT TAT AAC TATAAGG TAGAAGATTC ACCTCTGGTT TGCAATT.GCA GTGTTCATGA AGCCAAACTC AACGACACTC GAGTAATTTT CCAGTCACCT AAGCCTGATC CACCATTAGO TTTAAAGATT TCTTGCTCCA ATCAAGTGAA ATATTCAGAG AAGATTGTCT CAGCTACATC TTCGTATGAG GTTCAGGTGA GGAGTGACTG GAGTACTCCT TTTCCACCTA AAATTCTGAC CATCTATAAG AAGGAAAACA GGATGAATTT AGCTGAGAAA GATCATGTTA GCAAAGTTAC AGGAAAGTTT ACCTATGATG AACC TAT GAO

ATTCGAATGG

GGTACTCACT

GAGTGAGCAA

AGACATTTGT

AAC TGGAACA

TTATGTGGAG

TCCATCTTTT

CCCCAAAAAG

ATGTTGTGAA

TCCTTATGTG

CTAATGTCAG

TTTGCATATG

GCCCACCATT

AATTCTACAA

CCTGCTAGTA

GGGGCAAGAG

CGTGTCTTTA

AAGTGTTGGG

AGATTGTTCC

ATTCCTCAAA

TTTTTTCAAT

CAGTGTACTG

GACCCOCGGA TCAAGGTGTA TTCTGTGTGG TTTTGTTACA TAACTTGTCA TATCCAATTA 70 0** 1301 1351 1401 1451 1501 1551 1601 1651 1701 1751 1801 1851 1901 1951 30 2001 2051 2101 2151 2201 40 2251 2301 2351 2401 2451 2501 2551

CTGCAATGAA

ATGTCAATAT

ACTTGCAGAT

GCAATTGAGG

TTCATCCCAT

TATGAATGCA

GATTAGGATC

TCCT TCC TGA

GAAATTACTA

CTTTCCAGAG

AAGAAGTACA

GTCAGTCTCC

CTGTAAGAGG

CCTACACAGT

TGGAGAATAA

ACTTTGGAAG

ATGTGATAAA

GGAAATCACA

TACGGTTCTG

TAACCTTTTC

GCTTATCCTT

CAGTGATTAC

AAGATGGTGA

TATATCCATG

CCCAATATTT

CTCAAGATGA

CATGAATGCC

CAATATCTCA

GGTCAACCAG

TAT CAT AGGA AT CTGAGC CC

TTTTCCAGCC

AATCACTCTC

TTCTGTGGTG

TAAACATTGG

AATAAC CTTC

ATGGAAGATG

CAGTTCCAGA

CTAGATGGAC

TGTCATGGAT

TTAATG GAGA

CCCCTGATGA

CCATCATACT

CGAAATTCAC

GCCATCAATT

ATGGCC TATG

TAAACAGCAG

AAGCTAATGT

AATAAAATGG

ATCATT TTAT

ATGGAAGGAG

TATTGAAAAA

ATCATCGCTA TGCTGAATTA

TATGTGATTG

TGTGAAACTG ATGGGTACTT

AACTAAAATG

TACAATCCAG TCACTTGCGG

AAAGCACTTT

GCAGCCTTTA CTGTTCTGAT

ATTCCATCTA

AAAGATTGCT ATTTGCAGAG

TGATGGTTTT

AATCTTCCTA TTATCTGGCT

ACACAATGTG

TAGGTTCACT TGACTCTCCA

CCAACATGTG

AAGCCACTGC CTCCATCCAG TGTGAAAGCA ATTATTGAAA ATATCTTGGG AAAAGCCAGT AATTCCAGAT TCGCTATGGT TTAAGTGGAA TATGAGGTTT ATGATGCAAA ATCAAAATCT CTTGTGTCCA GTCTATGCTG TTCAGGTGCG TOGGATATTO GAGTAATTGG AGCAATCCAG ATAAAAGTTC CTATGAGAGG ACCTGAATTT TACTATGAAA AAGGAGAAAA ATGTCACTTT AAAATGACTC ATTGTGCAGT GTTCAGAGAT TCCTGCAATG GAACATGGTC AGAAGATGTG TTTCCTGTGG ACAGAGCAAG CACATACTGT CAATTGGTGC TTCTGTTGCA AATTTTAATT AGCAAAGTAA ATATCGTGCA GTCACTCAGT TTGTGTGATT GTTTCCTGGA TACTATCACC ATTTTATTAT TGAGTGGAAA AATCTTAATG CTTAGAATCT CTTCATCTGT TAAGAAGTAT CCCCATTGAG AAGTACCAGT TCAGTCTTTA TGGGAAAACC AAAGATAATT AATAGTTTCA CACCAGAGTG ATGCAGGTTT ATATGTAATT 71 2601 2651 2701 2751 2801 2851 2901 2951 3001 3051 GTGCCAGTA.A TTATTTCCTC TTCCATCTTA ATCACACCAA AGAATGAAAA AGCTATTTTG AGAATTGTTC CTGGGCACAA GGACTTAATT ATCTTTTTAT CAAGCATACA GCATCAGTGA GAGCCTGAAA CAATTTCAGA AGATATCAGT TAAAGATGAG ATGATCCCAA CAACTGTGGT ATCTTGAAAA GGGTTCTGTT TGTTTTAGTG TTCTCTGAGG CTGAGGGTAC TGAGGTAACC ACAACCCTTT GTTAAATACG CCACGCTGAT AAACTGGTGA AGA TTGCTTOGAA. CATTATTAAT GGAAGATGTT CCGAACCCCA TTCAGAAGAA ACGTTTGAGC CATGTGGTCC TCTTCTTTTG GTTGATACAT CATGCAAAAA CTCTCTACTT TCAACAACAG ACCAGTTCAA CAGTGTTAAC TATGAGGACG AAAGCCAGAG CAGCAACTCT AAACCAAGTG 72 Human OB Receotor Amino Acid Secruence (Sea, ID No. 5 (Amino 1 101 151 201 251 301 351 401 451 501 551 601 651 701 751 801 851 901 951 MI CQKFCVVL AGLSKNT SNS LCADN IEGKT

LFRNYNYKVH

PTAKLNDTLL

GNLKI SWSSP GS SYEVQVRG

HCIYKKENKI

PRGKFTYDAV

TSTIQSLAES

QP IFLLSGYT

IGLLKISWEK

PDLCAVYAVQ

GDTMKKEKNV

FTFLWTEQAHi

SSCVIVSWIL

Fl? IEKYQFS

SSSILLLGTL

SHHHSLISST

PS VRNTQE* S

LHWEFIYVIT

NGHYETAVEP

FVSTV'NS LVF LLYVLP EVLE MCLK ITS GGV P LVP FLQYQ KRLDGPG IWS VPSKE IVWWM

YCCNEHECHH

TLQLRYHRS S

MWIRINHSLG

PVEPENNLQF

VRCKRLDGLG

T LLWKP LMKN TVTVLAINS I SF SD YKLMYF LYP IFMEGVG

LISHQP.MKKL

QGHKH-CGRP Q I KKKKKKLE G AFNLSYPITP WRFKLSCMPP NSTYDYFLLP

KFNSSGTHFS

QQIDANWNIQ

DSP LVPQKGS

IFQSPLMSVQ

VKYSENSTTV

DWS TP RVF TT

NLAEKIPQSQ

RYAELYVI DV

LYCSDIPSIH

NLSKTTFHCC

CWLKGDLKLF

FQMVHCNC SV P INMVKPDPP

IREADKIVSA

QDVIYFPPKI

YDVVSDHVSK

NINISCETDG

PISEPKDCYL

FRSEQDRNCS

ICYVESLFKN

HECCECLVPV

LGLHME ITDD

TSLLVDSILP

LT SVG SNVS F

VTFFNLNETK

YLTKMTCRWS

QSDGFYECIF

SSVKAEITIN

AKSKSVSLPV

RGPEFWRI IN

WSEDVGNHTK

VQSLSAYPLN

SVKKYYIH-DH

GLYVIVPVI I

KMLEGSMFVK,

PLLSYDPAKS

so.

0 SLDSPPTCVL PDSVVKPLPP QIRYGLSGKE VQWKMYEVYD YWSNWSNPAY TVVMDIKVPM DSLCSVQRYV INHHTSCNGT GASVANFNLT FSWPMSKVNI I IEWKNLNED GEIKWLRISS KPKIINSFTQ DDIEKHQSDA FWEDVPNPKN CSWAQGLNFQ GPLHRKTRDL CSLVYLLTLP 73 Human OB Receotor "C"I DNA Seqruence (Se-aID No. 6(DNA): 2.

51 101 .0 151 201 251 301 351 ~0 401 451.

501 551 601 651 701 751 35 8021 851 901 951 1001 1051 1101 1151 1201

CCGCCGCCAT

CTTC TCTGAA

TTGGGAATTT

CTCCTTGGAG

TACTTCCTTT

ACATTATGAG

TTTCTAACTT

GATAGAAACT

TTCAACAGTA

TACAGTGCTG

TCATTATTTA

ATATGTTCTG

GCAGTTTTCA

TGTCTTGTGC

TTTGAAAATC

TTCAGCCCAT

GAAATCACAG

GGTACCATTT

CAGT TAT CAG

GACAGTATAC

ACTGGATGGC

CCACACAAGA

TCTAATGTTT

CTCAAAAGAG

GCCAGTATGA

CTCTGCCTTC

GTAAGATGAT

ATTTATGTGA

ATTTAAGTTG

TGCCTGCTGG

ACAGCTGTTG

ATCCAAAACA

GCTCCTTATG

AATTCTTTAG

GCTAAAAGGA

AGAATCTATT

CCTGAAGTGT

GATGGTTCAC

CTGTGCCAAC

ACATCTGGTG

AAATATGGTG

ATGATGGTAA

CCACTTCAAT

AGAAGCTGAC

TTCCTGGGTC

CCAGGAATCT

TGTCATATAC

CTTTTCACTG

ATTGTTTGGT

TGTTGTGAGT

GGTCGAGTTG

GACCCGGA

TTGTCAAAAA

TTCTGTGTGG

TAACTGCGTT

TAACTTGTCA

TCTTGCATGC CACCAAATTC ACTCTCAAAG AATACTTCAA AACCTAAGTT

TAATTCAAGT

ACTTTCCACT

GTTGCTTTCG

TGCAGACAAC ATTGAAGGAA TTTTTCAACA

AATAGATGCA

GACTTAAAAT TATTCATCTG CAGGAAT TAT AACTATAAGG TAGAAGATTC ACCTCTGGTT TGCAATTG3CA

GTGTTCATGA

AGCCAAACTC AACGACACTC GAGTAATTTT CCAGTCACCT AAGCCTGATC CACCATTAGG TTTAAAGATT TCTTGGTCCA ATCAAGTGAA ATATTCAGAG AAGATTGTCT CAGCTACATC TTCGTATGAG GTTCAGGTGA GGAGTGACTG GAGTACTCCT TTTCCACCTA AAATTCTGAC CATCTATAAG AAGGAAAACA GGATGAATTT AGCTGAGAAA GATCATGTTA GCAAAGTTAC

TCAAGGTGTA

TTTTGTTACA

TATCCAATTA

AACC TAT GAO

ATTCGAATGG

GGTACTCACT

GAGTGAGCAA

AGACATTTGT

AACTGGAACA

TTATGTGGAG

TCCATCTTT T CCC CAAAAAG

ATGTTGTGAA

TCCTTATGTG

CTAATGTCAG

TTTGCATATG

GCCCACCATT

AATTCTACAA

CCTGCTAGTA

GGGGCAAGAG

CGTGTCTTTA

AAGTGTTGGG

AGATTGTTC

AT TOCT CAAA

TTTTTTCAAT

74 1251 1301 1351 1401 1451 .0 1501 1551 1601 1651 1701 1751 1801 1851 1901 1951 2001 2051 35 2101 2151 2201 2251 2301 2351 2401 2451 )0 2501 2551

CTGAATGAAA

CTGCAATGAA

ATGTCAATAT

ACTTGCAGAT

GCAATTGAGG

TTCATCCCAT

TATGAATGCA

GATTAGGATC

TOCT TCCTGA

GAAATTACTA

CTTTCCACAG

AAGAAGTACA

GTCAGTCTCC

CTGTAAGAGG

CCTACACAGT

TGGAGAATAA

ACTTTGGAAG

ATGTGATAAA

GGAAATCACA

TACGGTTCTG

TAACCTTTTC

GCTTATCCTT

CAGTGATTAC

AAGATGGTGA

TATATCCATG

CCCAATATTT

CTCAAGATGA

CCAAACCTCG

CATGAATCC

CAATATCTCA

GGTCAACCAG

TATCATAGGA

ATCTGAGCCC

TTTTCCAGCC

AATCACTCTC

TTCTGTGGTG

TAAACATTGG

AATAACCTTC

ATGGAAGATG

CAGTTCCAGA

CTAGATGGAC

TGTCATGGAT

TTAATGGAGA

CCCCTGATGA

CCATCATACT

CGAAATTCAC

GCCATCAATT

ATGGCCTATG

TAAACAGCAG

AAGCTAATGT

AATAAAATGG

ATCATTTTAT

ATGGAAGGAG

TAT TGAAAAA

AGGA-AAGTTT

ATOATOOCTA

TGTGAAACTG

TACAATCCAG

GCAGCCTTTA

AAAGATTGCT

AATCTTCCTA

TAGG TTCACT

AAGCCACTGC

ATTATTGAAA

AATTCCAGAT

TATGAGGTTT

C TTGTGTGCA

TGGGATATTG

ATAAAAGTTC

TACTATGAAA

AAAATGACTC

TCCTGCAATG

TTTCCTGTGG

CAATTGGTGC

AGCAAAGTAA

TTGTGTGATT

ATTTTATTAT

CTTAGAATCT

CCCCATTGAG

T GGGAAAACC

CACCAGAGTG

ACCTATGATG

TOO TGAATTA ATOGGTACT T

TCACTTGCGG

CTGTTCTGAT

AT T TCAGAG

TTATCTGGCT

TGACTCTCCA

CTCCATCCAG

ATATCTTGGG

TCGCTATGGT

ATGATGCAAA

GTCTATGCTG

GAGTAATTGG

CTATGAGAGG

AAGGAGAAAA

ATTGTGCAGT

GAACATGGT C

ACAGAGCAAG

TTCTGTTGCA

ATATCGTGCA

GTTTCCTGGA

TGAGTGGAAA

CTTCATCTGT

AAGTACCAGT

AAAGATAAT T

ATGCAGGTTT

CAGTGTACTG

TATGTGATTG

AACTAAAATG

AAAGCACTTT

ATTCCATCTA

TGATGGTTTT

ACACAATGTG

CCAACATGTG

TGTGAAAGCA

AAAAGCCAGT

TTAAGTGGAA

ATCAAAATCT

TTCAGGTGCG

AGCAATCCAG

ACCTGAATTT

ATGTCACTTT

GTTCAGAGAT

AGAAGATGTG

CACATACTGT

AATT TTAATT

GTCACTCAGT

TACTATCACC

AATCTTAATG

TAAGAAGTAT

TCAGTCTTTA

AATAGTTTCA

ATATGTAATT

75 2601.

2651 2701 2751 LO0 2801 2851 2901 L 5 2951 GTGCCAGTAA TTATTTCCTO TTCCATCTTA TTGCTTGGAA ATCACACCAA AGAATGAAAA AGCTATTTTG CGAAGATGTT AGAATTGTTC CTGGGCACAA GGACTTAATT TTCAGAAGAT AGCATGTTCG TTAAGAGTCA TCACCACTCC CTAATCTCAA ACACAAACAC TGCGGAAGGC CACAGGGTCC TCTGCATAGG ACCTTTGTTC ACTTGTTTAT CTGCTCACCC TCCCTCCACT GACCCTGCCA AATCCCCCTC TGTGAGAAAC ACCCAACAAT AAAAAA AAAAAACTCC AGGGGG CAT TAT TAAT

CCGAACOCCA

GCTTGAAGGC

GTACCCAGGG

AAAACCAGAG

ATTGTCCTAT

GATCAATAAA

76 Human OB Recector I'D" Amino Acid Secmence (Secrience ID No. 7) 1 MICQKFCVVL LHWEFIYVIT AFNLSYPITP WRFKLSCI-2P 51 101 0 151 201 AGLSKNTSNS NGHYETAVEP LCADNIEGKT EVSTVNSLVF LFRNYNYKVH LLYVLP EVLE, PTAKLNDTLL MCLKITSGGV 251 301 351 ~0 401 451 501 551 ***.601 701 .*.*751 801 :851 951 1001 1051 1101

GNLKISWSSP'

GSSYEVQVRG

HCIYKKENKI

PRGKFTYDAV

TSTIQSLAES

QPIFLLSGYT

IGLLKI SWEK PD LCAVYAVQ

GDTMKKEKNV

FTFLWTEQAH

SSCVIVSWIL

FIPIEKYQFS

SSSILLLGTL

KIITASVTCGP

GSVCISDQFN

EQGLINSSVT

LTFSEGLDEL

SCPFPAPCLF

P LVPFP LQYQ

KRLDGPGIWS

VPSKE IVWWM

YCCNEHECHH

TLQLRYHRSS

MWIRINHSLG

PVFPENNLQF

VRCKRLDGLG

TLLWKP LMKN TVTVLAINS I SP SDYKLMYF LYP IFMEGVG

LISHQRMKKL

LLLEPETISE

SVNFSEAEGT

KCFSSKNSPL

LKLEGNFPEE

TO IRVLQD SC KFNSSGTHFS NLSKTTFHCC QQIDANWNIQ CWLKGDLKLF DSPLVPQKGS FQMVHCNCSV IFQSPLMSVQ PINMVKPDPP VKYSENSTTV IREADKIVSA DWSTPRVFTT QDVIYFPPKI NLAEKIPQSQ YDVVSDHVSK RYAELYVIDV NINISCETDG LYCSDIPSIH PISEPKDCYL SLDSPPTCVL PDSVVKPLPP QIRYGLSGKE VQWKMYEVYD YWSNWSNPAkY TVV1M IKVPM

NSTYDYFLLP

FRSEQDRNCS

ICYVESLFKN

HECCECLVPV

LGLHME ITDD

TSLLVDSILP

LTSVGSNVSF

VTFFNLNETK

YLTKMTCRWS

QSDGFYECIF

SSVKAEITIN

AKSKSVSLPV

RGPEFWRI IN

WSEDVGNHTK

VQSLSAYPLN

SVKKYYIHDH

GLYVIVPVI I KPETFEHLF I

SLLSTTDLEK

SNSKP SETGE

DQHPNIISPH

GVLLTDKSRV

YMPQFQTCST

DSLCSVQRYV

GAS VANFNLT I IEWKNLNED

KPKIINSFTQ

FWEDVPNPKN

DISVDTSWKN

EVTYEDESQR

KDSFSNS SWE NNDKKS IYYL

SHFVENNINL

INHHTSCNGT

FSWPMSKVNI

GEIKWLRISS

DDIEKHQSDA

CSWAQGLNFQ

KDEMMPTTVV

QPFVKYATL I

IEAQAFFILS

GVT SI KKRE S GTS SKKTFAS )0 1151 1201 QTHKIMENKM CDLTV*FH*R ENNCSK*KKK KKNSRPARPD NLQICVIMGN IKCNRL*LWV GERKETRVKF 77 Human OB Receipt-or Nucleic Acid Secruence(Seqruence ID No. 8) 1 51 101.

151 0 201 251 301 351 401 451 501 551 601 651.

701 751 15 801 851 901 951 1001 1051 110~1 1151 io 1201 1251

GCGGCCGCCA

GGTCGAGTTG

TTGTCAAAAA

TAACTGCGTT

TCTTGCATGC

GCTCTCAAAG

AACCTAAGTT

ACTTTCCACT

TGCAGACAAC

TTTTTCAACA

GACT TAAAAT

CAGGAATTAT

TAGAAGATTC

TGCAATTGCA

AGCCAAACTC

GAGTAATTTT

AAGCCTGATC

TTTAAAGATT

ATCAAGTGAA

AAGATTGTCT

TTCGTATGAG

GGAGTGACTG

TTTCCACCTA

CAT CTATAAG

GGATGAATTT

GATCATGTTA

GTGTGATGGA

GACCCCCGGA

TTCTGTGTGG

TAACTTGTCA

CACCAAATTC

AATACT TCAA

TAATTCAAGT

GTTGCTTTCG

ATTGAAGGAA.

AATAGATGCA

TATTCATCTG

AACTATAAGG

ACCT C TGGT T

GTGTTCACGA

AACGACACTC

CCAGTCACCT

CAC CAT TAGG

TCTTGGTCCA

ATATTCAGAG

CAGCTACATC

GTTCAGGTGA

GAGTACTCCT

AAATTCTGAC

AAGGAAAACA

AGCTGAGAAA

GCAAAGTTAC

TATCTGCAGA

TCAAGGTGTA

TTTTGTTACA

TATCCAATTA

AACCTATGAC

ATTCGAATGG

GGTACTCACT

GAGTGAGCAA

AGACATTTGT

AACTGGAACA

TTATGTGGAG

TCCATCTTTT

CCCCAAAAAG

ATGTTGT GAA

TCCTTATGTG

CTAATGTCAG

TTTGCATATG

GCCCACCATT

AATTCTACAA

CCTGCTAGTA

GGGGCAAGAG

C GT GTC TTTA

AAGTGTTGGG

AGATTGTTCC

ATTCCTCAAA

TTTTTTCAAT

ATTCGGCTTT

CTCTGCCTTC

CTTCTCTGAA

GTAAGATGAT

TTGGGAATTT

ATTTATGTGA

CTCCTTGGAG

ATTTAAGTTG

TACTTCCTTT

TGCCTGCTGG

ACATTATOAG

ACAGCTGTTG

TTTCTAACTT ATCCAAAACA GATAGAAACT GCTCCTTATG TTCAACAGTA AATTCTTTAG TACAGTGCTG GCTAAAAGGA TCATTATTTA AGAATCTATT

ATATGTTCTG

GCAGTTTTCA

TGTCTTGTGC

TTTGAAAATC

TTCAGCCCAT

GAAATCACAG

GGTACCATTT

CAGTTATCAG

GACAGTATAC

ACT GGATGGC

CCACACAAGA

TCTAATGTT T

CTCAAAAGAG

CCTGAAGTGT

GATGGTTCAC

CTGTGCCAAC

ACATCTGGTG

AAATATGGTG

ATGATGGTAA

CCACTTCAAT

AGAAGCTGAC

TTCCTGGGTC

CCAGGAATCT

TGTCATATAC

CTTTTCACTG

ATTGTTTGGT

GCCAGTATGA TGTTGTGAGT CTGAATGAAA CCAAACCTCG 78 1301 1351 1401 1451 L.0 1501 1551 1601 1651 1701 ~0 1751 1801 1851 1901 :1951 2001 2051 2101 .*..*2151 :2201.

1 0 2251 2301 2351 2401 2451 2501 2551

AGGAAAGTTT

ATCATCGCTA

TGTGAAACTG

TACAATCCAG

GCAGCCTTTA

AAAGATTGCT

AATCTTCCTA

TAGGTTCACT

AAGCCACTGC

ATTATTGAAA

AATTCCAGAT

TATGAGGTTT

CTTGTGTGCA

TGGGATATTG

ATAAAAGTTC

TACTATGAAA

AAAATGACTC

TCCTGCAATG

TTTCCTGTGG

CAATTGGTGC

AGCAAAGTAA

TTGTGTGATT

ATTTTATTAT

CTTAGAATCT

CCCCATTGAG

TGGGAAAACC

ACCTATGATG CAGTGTACTC CTGCAATGAA TGCTGAATTA TATGTGATTG ATGTCAATAT ATGGCTACTT AACTAAAATC ACTTGCAGAT TCACTTGCGG AAAGCACTTT CTGTTCTGAT ATTCCATCTA ATTTGCAGAG TGATGGTTTT TTATCTGGCT ACACAATGTG TGACTCTCCA CCAACATGTG CTCCATCCAG TGTGAAAGCA ATATCTTGGG AAAAGCCAGT TCGCTATGGT TTAAGTGGAA ATGATGCAAA ATCAAAATCT GTCTATCCTG TTCAGGTGCG GAGTAATTGG AGCAATCCAG CTATGAGAGG ACCTGAATTT AAGGAGAAAA ATGTCACTTT ATTGTGCAGT GTTCACAGAT GAACATGGTC AGAAGATGTG ACAGAGCAAG CACATACTGT TTCTGTTGCA AATTTTAATT ATATCGTGCA GTCACTCAGT GTTTCCTGGA TACTATCACC TGAGTGGAAA AATCTTAATG CTTCATCTGT TAAGAAGTAT AAGTACCAGT TCAGTCTTTA AAAGATAATT AATAGTTTCA

GCAATTGAGG

TTCATCCCAT

TATGAATGCA

GAT TAGGAT C

TCCTTCCTGA

GAAAT TACTA

CTTTCCAGAG

AAGAAGTACA

GTCAGTCTCC

CTG TAAGAGG

CCTACACAGT

TGGAGAATAA

ACT TTGGAAG

ATGTGATAAA

GGAAATCACA

TACGGTTCTG

TAACCTTTTC

GCTTATCCTT

CAGTGATTAC

AAGATGGTGA

TATATCCAT G

CCCAATATTT

CTCAAGATGA

CATGAATGCC

CAATATCTCA

GGTCAACCAG

TATOATAGGA

ATCTGAGCCC

TTTTCCAGCC

AATCACTCTC

TLTCTGTGGTG

TAAACATTGG

AATAACCTTC

ATGGAAGATG

CAGTTCCAGA

CTAGATGGAC

TGTCATGGAT

TTAATGGAGA

CCCC TGATGA

CCATCATACT

CGAAATTCAC

GCCATCAATT

ATGGCCTATG

TAAACAGCAG

AAGCTAATGT

AATAA.AATGG

ATCATTTTAT

ATGGAAGGAG

TATTGAAAAA

79 2601 2651 2702.

2751 2801 2851 2902.

2951 3002.

3051 3101 3151 25 3201 3251 3301 3351 3401 35 3451 3501

CACCAGAGTG

TTCCATCTTA

AGCTATTTTG

GGACTTAATT

TACAGCATCA

CAGAAGATAT

CCAACAACTG

TGTTTGTATT

GTACTGAGGT

TACGCCACGC

AGGGCTTATA

CGTTGAAGGA

TTTTTTATAT

ATTCTCAGAA

AAGAAAATAA

AAAAAGAGAG

CCCATTCCCA

GTTGCTCACA

AAGACTTTTG

ATGCAGGTTT

TTGC TTGGAA

GGAAGATGTT

TTCAGAAGCC

GTGACATGTG

CAGTGTTGAT

TGGTCTCTCT

AGTGACCAGT

AACCTATGAG

TGATCAGCAA

AATAGT TCAG

TTCTTTCTCT

TATCGGATCA

GGATTGGATG

TGATAAAAAG

AGAGTGGTGT

GCCCCCTGTT

CTTTGTAGAA

CATCTTACAT

ATATGTAATT

CATTATTAAT

CCGAACCCCA

AGAAACGTTT

GTCCTCTTCT

ACATCATGGA

ACTTTCAACA

TCAACAGTGT

GACGAAAGCC

CTCTAAACCA

TCACCAAGTG

A.ATAGC TCAT

GCATCCCAAC

AACTTTTGAA

TCTATCTATT

GCTTTTGACT

TATTCACGGA

AATAATATCA

GCCTCAATTC

GTGCCAGTAA

ATCACACCAA

AGAATTGTTC

GAGCATCTTT

TTTGGAGCCT

AAAATAAAGA

ACAGATCTTG

TAACTTCTCT

AGAGACAACC

AGT GAAAC TG

CTTCTCTAGC

GGGAGATAGA

ATAATTTCAC

ATTGGAGGGA

ATT TAGGGG T

GACAAGTCAA

CATCAGAGTT

ACTTAGGAAC

CAAACTTGT T

TTATTTCCTC

AGAATGAAAA

CTGGGCACAA

TTATCAAGCA

GAAACAATTT

TGAGATGATG

AAAAGGGTTC

GAGGCC TGAGG

CTTTGTTAAA

GTGAAGAACA

AAAAATTCTC

GGCCCAGGCA

CACACCTCAC

AATTTCCCTG

CACC TCAATC

GGGTATCGTG

CTCCAGGACA

TTCTAGTAAG

CTACTCAGAC

4@e* St..

S. 55 S S

S

S.

S S

S.

S

9

S

5* S Se

S.

S 3551 TCATAAGATC ATGGAAAACA AGATGTGTGA CCTAACTGTG TAATCTAGA 80 Human OB Receptor Protein Chromosomal DNA (Sec. ID No. 9) 0 @000

S

S.

*6

S

S

I

S.

0 *50* -0 CAT TGG G gtaagttatttg.....

His Trp Glu 12 13 14 CAA ATA G gtaagcattagc Gin Ile Asp 122 123 124 TAT GTT CT gtaagtaccaaa.....

Tyr Val Leu 163 164 165 AAT ATG G gtaagttatgca.....

Asn Met Val 233 234 235 ATC AGA GAA gtaagtatattt.....

Ile Arg Glu 281 282 283 ACA CAA G gtaggttatgta.....

Thr Gin Asp 330 331 332 GTG ATT G gtaagaaaacag.....

Val Ile Asp 427 428 429 TAT CAT AG gtacgtattatt.....

Tyr His Arg 466 467 468 TCT GTG G gtatgtcaagct.....

Ser Vai Val 533 534 535 CAA TGG AAG gtaccttttact.....

Gin Trp Lys 582 583 584 ATA AAA G gtctgagagat.....

Ile Lys Val 636 637 638 Intron 1 taccttttccag Intron 2 atatcctaacag Intron 3 ttttaaattcag Intron 4 Intron 5 Intron 6 Intron 7 Intron 8 ttttcaatatag tttttccttaag aatatttaacag ccctcattacag tgtttcaaatag AA TTT ATT Phe Ile 15 16 AT GCA AAC Ala Asn 125 126 G CCT GAA Pro Glu 166 167 TG AAG CCT Lys Pro 236 237 GCT GAC AAG Ala Asp Lys 284 285 286 AT GrC ATA Val Ile 333 334 AT GTC AAT Val Asn 430 431 G AGC AGC Ser Ser 469 470 TG AG CCA Lys Pro 536 537 ATG TAT GAG Met Tyr Glu 585 586 587 TT CCT ATG Pro Met 639 640 GTG TAC TTC Intron 9 tatcttttaaag Intron 10 aaaaatttctag Intron 11 cttattttacag Intron 12 gtcattttgcag 81 CTr TGG Leu Trp 663 664 AGC AAA Ser Lys 0 736 737 ATC CAT Ile His 797 798 ACT CAA Thr Gin '0 829 830 CAC CAA His Gin 864 865 iri CAG Phe Gin 0 889 890 AAG gtattcccaatt.....

Lys 665 G gtaagaagaggt.....

Vai 738 G gtaagtttacta.....

Intron 13 tatttacacag Intron 14 ttttcccctcag Intron 15 ttttctcctcag G gtaaaaattata..... Intron 16 tttctttttcag AG gtattgtacttg.....

Arg 866 AAG gttgctttttca.....

Lys 891 Intron 17 tatcctttgtag Intron 18 ttatctaaacag CCC CTG ATG Pro Leu Met 666 667 668 TA AAT ATC Asn Ile 739 740 AT CAT TTI His Phe 800 801 AT GAT ATT Asp Ile 832 833 A ATG AAA Met Lys 867 868 Exon A AGA ACG GAC Arg Thr Asp 892 893 894 Exon 0 CCA GAA ACG Pro Giu Thr 892 893 894 Exon B AAA CGT TTG Lys Arg Leu 892 893 894 Exon C ATG CTr GAA Met Ieu Glu 892 893 894 0* Exon A AAA TAT GAT gtacatttgtct..... Iniron 18 cttttcttttag Exon D GAA ACC AGA gtatccagtgtt..... Intron 18 ctttttaaacag 82 Human OB Receiptor Protein, Recombinant Secreted-Raee-tor amino acid seqruence (Se2. ID. No. 1 51 1021 151 201 251 301 351 401 451 501 551 601 651 701 751 MI CQKFCVVL

AGLSKNTSNS

LCADN IEGKT LFRN YNYKVH P TAKLNDTLL

GNLKISWSSP

GSSYEVQVRG

HCIYKKENKI

PRGKFTYDAV

TSTIQSLAES

QP IFLLSCYT

IGLLKISWEK

PDLCAVYAVQ

GDTMKKEKNV

FTFLWTEQAH

35 CV IVS WI L

LHWEFIYVIT

NGHYETAVEP

F VS TVN SLVF

LLY'VLPEVLE

MCLKITSGGV

P LVPFP LQYQ

KRLDGPGIWS

VPSKEIVWWM

YCCNEHECHH

TLQLRYHRSS

MWIRINHSLG

PVFPENNLQF

VRCKRLDGLG

TLLWKP LMKN TVTVLAINS I

SPSDYKLMYF

AFNLSYP ITP WRFKLSCMPP NSTYDYFLLp KFNSSGTHFS NLSKTTFHiCC FRSEQDp NCS QQIDANWNIQ CWLKGDLKLF ICYVESLFKN DSPLVPQKGS FQMVHCNCSV H-ECCECLVPV IFQSPLMSVQ ?INMVKPDPP LGLHIVEITDD VKYSENSTTV IREADKIVSA TSLLVDSILP DWSTPRVFTT QDVIYFPPKI LTSVGSNVSF NLAEKIPQSQ YDVVSDHVSK VTFFNLNETK RYAELYVIDV NINISCETDG YLTKMTCRWS LYCSDIPSIH PISEPKDCYL QSDGFYECIF SLDSPPTCVL PDSVVKPLPP SSVKAEITIN QIRYGLSGKE VQWKMYEVYD AKSKSVSLPV YWSNWSNPAY TVVMDIKVPM RGPEFWRIIN DSLCSVQRYV INHHTSCNGT WSEDVGNHTK GASVANFNLT FSWPMSKVNI VQSLSAYPLN IIEWKNLNED GEIKWLRISS SVKKYYIHDH 0 00 0 a a. 0 *0a.

801 FIPIEKYQFS LYPIFMEGVG KPKIINSFTQ DDIEKHQSD 83 Human 08 Receotor Protein. Recombinant Secreted Receptor DNA sequience (Seq. 1D. No. 11): 1.

51 101 0 152.

202.

251 301 351 0 401 451 501 551 601 651 701 751 801 851 t0 901 951 1001 1051 1101 1151 CGG CCGC CA

GGTCGAGTTG

TTGTCAAAAA

TAACTGCGTT

TCTTGCATC

GCTCTCAAAG

AACCTAAGTT

ACTTTCCACT

TGCAGACAAC

TTTTTCAACA

GACT TAAAAT

CAGGAATTAT

TAGAAGATTC

TGCAATTGCA

AGCCAAACTC

GAGTAATTTT

AAGCCTGATC

TTTAAAGATT

ATCAAGTGAA

AAGATTGTCT

TTCGTATGAG

GGAGTGACTG

TTTCCACCTA

CATC TATAAG

GTGTGATGGA

GAC CC C CGGA

TTCTGTGTGG

TAACTTGTCA

CACCAAATTC

AATACTTCAA

TAATTCAAGT

GTTGCTTTCG

AT TGAAGGAA

AATAGATGCA

TATTCATCTG

AACTATAAGG

ACCTCTGGTT

GTGTTCACGA

AACGACACTC

CCAGTCACCT

CACCAT TAGG

TCTTGGTCCA

ATATTCAGAG

CAGCTACATC

GTTCAGGTGA

GAGTACTCCT

AAATTCTGAC

TATCTGCAGA ATTCGGCTTT TCAAGGTGTA CTTCTCTGAA TTTTGTTACA TTGGGAATTT TATCCAATTA CTCCTTGGAG AACCTATGAC TACTTCCTTT ATTCGAATGG ACATTATOAG GGTACTCACT TTTCTAACTT GAGTGAGCAA CAT AGAAAC T AGACATTTGT TTCAACAGTA AACTGGAACA TACAGTGCTG TTATGTGGAG TCATTATTTA TCCATCTTTT ATATGTTCTG CCCCAAAAAG GCAGTTTTCA ATGTTGTGAA TGTCTTGTGC TCCTTATGTG TTTGAAAATC CTAATGTCAG TTCAGCCCAT TTTGCATATG GAAATCACAG GCCCACCATT GGTACCATTT AATTCTACAA CAGTTATCAG CCTGCTAGTA GACAGTATAC GGGGCAAGAG ACTGGATGGC CGTGTCTTTA CCACACAAGA AAGTGTTGGG TCTAATGTTT

CTCTGCCTTC

GTAAGATGAT

ATTTATGTGA

ATTTAAGTTG

TGCCTGCTGG

ACAGCTGTTG

ATCCAAAACA

GCTCCTTATG

AATTCTTTAG

GCTAAAAGGA

AGAATCTATT

CCTGAAGTGT

GATGGTTCAC

CTGT GCCAAC

ACATCTGGTG

AAATATGGTG

ATGATGGTAA

CCACTTCAAT

AGAAGCTGAC

TTCCTGGGTC

CCAGGAATCT

TGTCATATAC

CTTTTCACTG

ATTGTTTGGT

AAGGAAAACA AGATTGTTCC CTCAAAAGAG 1201 GGATGAATTT AGCTGAGAAA ATTCCTCAAA GCCAGTATGA TGTTGTGAGT 84 1251.

1301 1351 1401 0 1451 1501 1551 1601 1651 ~0 1701 1751 1801 1851 1901 30 1951 2001 2051 2101 2151 2201 ****2251 2301 2351 2401 2451 2501

GATCATGTTA

AGGAAAGTTT

ATCATCGCTA

TGTGAAACTG

TACAATCCAG

GCAGCCTTTA

AAAGATTGCT

AATCTTCCTA

TAGGTTCACT

AAGCCACTGC

ATTATTGAAA

AATTCCAGAT

TATGAGGTTT

CTTGTGTGCA

TGGGATATTG

ATAA.AAGTTC

TACTATGAAA

AAAATGACTC

TCCTGCAATG

TTTCCTGTGG

CAATTGGTGC

AGCAAAGTAA

TTGTGTGATT

ATTTTATTAT

CTTAGAATCT

CCCCATTGAG

GCAAAGTTAC

ACOTATGATG

TGCTGA.ATTA

ATGGGTACTT

TCACTTGCGG

CTGTTCTGAT

ATTTGCAGAG

TTATCTGGCT

TGACTCTCCA

CTCCATCCAG

ATATOT TGGG

TCGCTATGGT

ATGATGCAAA

GTCTATGCTG

GAGTAATTGG

CTATGAGAGG

AAGGAGAAAA

ATTGTGCAGT

GAACATGGTC

ACAGAGCAAG

TTCTGTTGCA

ATATCGTGCA

GTTTCCTGGA

TGAGTGGAAA

CTTCATCTGT

AAG;TACCAGT

TTTTTTCAAT CTGAATGAAA CAGTGTACTG CTGCAATGAA TATGTGATTG ATGTCAATAT AACTAAAATG ACTTGCAGAT AAAGCACTTT GCAATTGAGG ATTCCATCTA TTCATCCCAT TGATGGTTTT TATGAATGCA ACACAATGTG GATTACCATO CCAACATGTG TCCTTCCTGA TGTGAAAGCA GAAATTACTA AAAAGCCAGT CTTTCCAGAG TTAAGTGGAA AAGAAGTACA ATCAAAATCT GTCAGTCTCC TTCAGGTGCG CTGTAAGAGG AGCAATCCAG CCTACACAGT ACCTGAATTT TGGAGAATAA ATGTCACTTT ACTTTGGAAG GTTCAGAGAT ATGTGATAAA AGAAGATGTG GGAAATCACA CACATACTGT TACGGTTCTG AATTTTAATT TAACCTTTTC GTCACTCAGT GCTTATCCTT TACTATCACC CAGTGATTAC

OCAAACCTCG

CATGAATGCC

CAATATCTCA

GGTCAACCAG

TATCATAGGA

ATCTGAGCCC

TTTTCCAGCC

AATCACTCTC

TTCTGTGGTG

TAAACATTGG

AATAACCTTC

ATGGAAGATG

CAGT TOCAGA C TAGATGGAC

TGTCATGGAT

T TAATGGAGA

CCCCTGATGA

CCATCATACT

CGAAATTCAC

GCCATCAATT

ATGGCCTATG

TAAACAGCAG

AAGC TAATGT AATCTTAATG AAGATGGTGA AATAAAATGG TAAGAAGTAT TATATCCATG ATCATTTTAT TCAGTCTTTA CCCAATATTT ATGGAAGGAG 85 2551 TGGCAAAACC AAAGATAATT AATAOTTTCA CTCAACATOA TATTGAAAA~.

2601 CACCAGAGTG ATTGATAAGG ATCC

S

S.

S S

S.

S

S

5555

S

5555 5555 5S55 S S

SSSS

86 Human OB Recpntor Protein. Recomnbinant S~zreted prnentcr T)MA quenince with C-terminal FLAG (Seqa. TD. No. 12) 1 51 101 CCATTGAAGT CAA.TGGGAGT TTGTTTTGGC ACCAA.AATCA ACGGGGATTT CCAAAATGTC GTAATAACCC CGCCCCGTTG ACGCAAATGC GCGGTAGGCG TGTACGGTGG GAGGTCTATA TAAGCAGAGC TCGTTTAGTG AACCGTCAGA 151 TCTCTAGAAG CTGGGTACCA 201 251 301 351 401 451 501 551 601 651 701 751 801 851 901 951 1001 1051 1101 1151 TGTGATGGAT ATCTGCAGAA ACCCOGGAT CAAGGTGTAC TCTGTGTGGT TTTGTTACAT AACTTGTCAT ATCCAATTAC ACCAAATTCA ACCTATGACT ATACTTCAAA TTCGAATGGA APJTCAAGTG GTACTCACTT TTGCTTTCGG AGTGAGCAAG TTGAAGGAAA GACATTTGTT ATAGATGCAA ACTGGAACAT ATTCATCTGT TATGTGGAGT ACTATAAGGT CCATCTTTTA CCTCTGGTTC CCCAAAAAGG TGTTCACGAA TGTTGTGAAT ACGACACTCT CCTTATGTGT CAGTCACCTC TAATGTCAGT GCTGCTAGCA AGCTTGCTAG TTCGGCTTTC TOTGCCTTCG TTCTCTGAAG TAAGATGATT TGGGAATTTA TTTATGTGAT TCCTTGGAGA TTTAAGTTGT ACTTCCTTTT GCCTGCTGGG CATTATGAGA CAGCTGTTGA TTCTAACTTA TCCAAAACAA ATAGAAACTG CTCCTTATGT TCAACAGTAA ATTCTTTAGT ACAGTGCTGG CTAAAAGGAG CATTATTTAA GAATCTATTC TATGTTCTGC CTGAAGTGTT CAGTTTTCAG ATGGTTCACT

CGGCCGCCAG

GTCGAGT TGG

TGTCAAAA.AT

AACTGCGTTT

CTTGCATGCC

o TCTCAAAGA

ACCTAAGTTT

CTTTCCACTG

GCAGACAACA

TTTTCAACAA

ACTTAAAATT

AGGAATTATA

AGAAGATTCA

GCAATTGCAG

GTCTTGTGCC TGTGCCAACA GCCAAACTCA TTGAAAATCA CATCTGGTGG AGTAATTTTC TCAGCCCATA AATATGGTGA AGCCTGATCC

ACCATTAGGT

CTTGGTCCAG

TATTCAGAGA

AGCTACATCC

TTGCATATGG AAATCACAGA TGATGGTAAT TTAAAGATTT CCCACCATTG GTACCATTTC CACTTCAATA TCAAGTGAAA ATTCTACAAC AGTTATCAGA GAAGCTGACA AGATTGTCTC CTGCTAGTAG ACAGTATACT TCCTGGGTCT TCGTATGAGG 87

S

S

12021 1251 1301 1351 1401 1451 1501 1551 1601 1651 1701 1751 1801 1851 1901 1951 2001 2051 2101 2151 2201 2251 2301 2351 2401 2451 2501

TTCAGGTGAG

AGTACTCCTC

AATTCTGACA

AGGAAAACAA

GCTGAGAAAA

CAAAGTTACT

CCTATGATGC

GCTGAAT TAT TGGG TACT TA

CACTTGCGGA

TGTTCTGATA

TTTGCAGAGT

TATCTGGCTA

GACTCTCCAC

TCCATCCAGT

TATCTTGGGA

CGCTATGGTT

TGAT GCAAAA

TCTATGCTGT

AGTAATTGGA

TATGAGAGGA

AGGAGAAAAA

TTGTGCAGTG

AACATGGTCA

CAGAGCAAGC

TCTGTTGCAA

TATCGTGCAG

GGGCAAGAGA

GTGTCTTTAC

AGTGTTGGGT

GATTGTTCCC

TTCCTCAAAG

TTTTTCAATC

AGTGTACTGC

ATGTGATTGA

ACTAAAATGA

AAGCACTTTG

TTCCATCTAT

GATGGTTTTT

CACAATGTGG

CAACATGTGT

GTGAAAGCAG

AAAGCCAGTC

TAAGTGGAAA

TCAAAATCTG

TCAGGTGCGC

GCAATCCAGC

CCTGAATTTT

TGTCACTTTA

TTCAGAGATA

GAAGATGTGG

ACATACTGTT

ATTTTAATTT

TCACTCAGTG

CTGGATGGCC CAGGAATCTG GAGTGACTGG CACACAAGAT GTCATATACT TTCCACCTAA CTAATGTTTC TTTTCACTGC ATCTATAAGA TCAAAAGAGA TTGTTTGGTG GATGAATTTA CCAGTATGAT GTTGTGAGTG ATCATGTTAG TGAATGAAAC CAAACCTCGA GGAAAGTTTA TGCAATGAAC ATGAATGCCA TCATCGCTAT TGTCAATATC AATATCTCAT GTGAAACTGA CTTGCAGATG GTCAACCAGT ACAATCCAGT CAATTGAGGT ATCATAGGAG CAGCCTTTAC TCATCCCATA TCTGAGCCCA AAGATTGCTA ATGAATGCAT TTTCCAGCCA ATOTTOCTAT ATTAGGATCA ATCACTCTCT ACGTTCACTT CCTTCCTGAT TCTGTGGTGA AGCCACTGCC AAATTACTAT AAACATTGGA TTATTGAAAA TTTCCAGAGA ATAACCTTCA ATTCCAGATT AGAAGTACAA TGGAAGATGT ATGAGGTTTA TCAGTCTCCC AGTTCCAGAC TTGTGTGCAG TGTAAGAGGC TAGATGGACT GGGATATTGG CTACACAGTT GTCATGGATA TAAAAGTTCC GGAGAATAAT TAATGGAGAT ACTATGAAAA CTTTGGAAGC CCCTGATGAA AAATGACTCA TGTGATAAAC CATCATACTT CCTGCAATGG GAAATCACAC GAAATTCACT TTCCTGTGGA ACGGTTCTGG CCATCAATTC AATTGGTGCT AACCTTTTCA TGGCCTATGA GCAA.AGTAAA CTTATCCTTT AAACAGCAGT TGTGTGATTG 88 2551 2601 2651 2701 .0 2751 2801 2851 2901

TTTCCTGGAT

GAGTGGAAAA

TTCATCTGTT

AGTACCAGTT

AAGATAAT TA

TGCAGGTGAC

AGATACATTG

ATGC TTTATT ACTATOACCO AGTGATTACA AGCTAATGTA TTTTATTATT ATCTTAATOA ACATGGTGAA ATAAAATGGC TTAGAATCTC AAGAAGTATT ATATCCATGA TCATTTTATC CCCATTGAGA CAOTCTTTAC CCAATATTTA TGGAAGGAGT GGGAAAACCA ATAGTTTCAC TCAAGATGAT ATTGAA.AAAC ACCAGAGTGA TACAAGGACG ACGATGACAA GTAGGGATCC AGACATGATA ATGAGTTTGG ACAACCCACA.ACTAGAATGC AGTGAAAAAA TGTGAAATTT GTGATGCTAT TOCTTTATTT GTAACCAT a.

a 89 Recombinant Human OB Receptor Protein,. Natural SpliceQ Variant amino acid secruence (Sear. ID. No. 13) 1 51 101.

151 201 251 301 351 401 451 501 551 601 30 651 701 751 .:801 MI CQKFCVVL AGLS KNT SN S

LCADNIEGKT

LFRNYNYKVH

P TAKLNDTLL GNLKI SWSSP GS SYEVQVRG

HCIYKKENKI

PRGKFTYDAV

TSTIQSLAES

QPIFLLSGYT

IGLLKISWEK

PDLCAVYAVQ

GD TMKKEKNV

FTFLWTEQAH

SSCVIVSWIL

FTIL

LH-WEFIYVIT

NGHYETAVEP

FVS TVNS LVF LLYVLP EVLE

MCLKITSGGV

PLVPFPLQYQ

-KRLDGPGIWS

VP SKE IVWWM

YCCNEFIECHH

TLQLRYH-RS S

MWIRINHSLG

PVFPENNLQF

VRCKRLDGLG

TLLWKPLMKN

TVTVLAINS I SF SDYKLMYF AFNLSYP ITP KFNS SGTHF S

QQIDANWNIQ

DSP LV? QKG S

IFQSPLMSVQ

VKYSENSTTV

DW STP RVF TT

NLAEKIPQSQ

RYAELYVI DV LYCSDIPS IH

SLDSPPTCVL

QIRYGLSGKE

YWSNWSNPAY

DSLCSVQRYV

WRFKLSCPP

NLSKTTFHCC

CWLKGDLKLF

FQMVHCNC SV

PINMVKPDPP

IREADKIVSA

QDVIY-FPPKI

YDVVS DHVS K NINI SCETDG PT SEPKDCYL

PDSVVKPLPP

VQWKMYEVYD

TVVMD IKVPM

INHHTSCNGT

NSTYDYFLLP

FRSEQDRNCS

ICYVESLFKN

HECCECLVPV

LGLHMEITDD

TSLLVDSILP

LTSVGSNVSF

VTFFN LNETK

YLTKMTCRWS

QSDGFYECIF

SSVKAEITIN

AKSKSVSLPV

RGPEFWRI IN

WSEDVGNHTK

VQSLSAYPLN

SVKKYYIHGK

GASVANFNLT FSWPMSKVNI I IEWKNLNED GEIKWLRISS *0 90 Human OB Receotor Protein, Natural S;plice variant DNA -(Seca. ID, No. 14) 102.

0 2151 201 251 301 351 0 402.

451 501 551 601 0 651 701 751 801 851 0 901 951 1001 1051 1101 .0 1151 1201

GCGGCCGCCA

GGTCGAGTTG

TTGTCAAAAA

TAACTGCGTT

TCTTGCATGC

GCTCTCAAAG

AACCTAAGTT

ACTTTCCACT

TGCAGACAAC

TTTTTCAACA

GACT TAAAAT CAGGAAT TAT

TAGAAGATTC

TGCAATTGCA

AGCCAAACTC

GAGTAATTTT

AAGCCTGATC

TTTAAAGATT

ATCAAGTGAA

AAGATTGTCT

TTCGTATGAG

GGAGTGACTG

T TTCCACCTA

GTGTGATGGA

GACCCCCGGA

TTCTGTGTGG

TAACTTGTCA

CACCAAATTC

AATACTTCAA

TAATTCAAGT

GTTGCTTTCG

AT TGAACGAA

AATAGATGCA

TATTCATCTG

AACTATAAGG

ACCTCT GGTT

GTGTTCACGA

AACGACACTC

CCAGTCACCT

CACCATTAGG

TCTTGGTCCA

ATATTCAGAG

CAGCTACATC

GTTCAGGTGA

GAGTACTCCT

AAATTCTGAC

TATCTGCAGA ATTCGGCTTT TCAAGGTGTA CTTCTCTGAA TTTTGTTACA TTGGGAATTT TATCCAATTA CTCCTTGGAG AACCTATGAC TACTTCCTTT ATTCGAATGG ACATTATGAG GGTACTCACT TTTCTAACTT GAG TGAG CAA GATAGAAAC T AGACATTTGT TTCAACAGTA AACTGGAACA TACAGTGCTG TTATGTGGAG TCATTATTTA TCCATCTTTT ATATGTTCTG CCCCAAAAAG GCAGTTTTCA ATGTTGTGAA TGTCTTGTGC TCCTTATGTG TTTGAAAATC CTAATGTCAG TTCAGCCCAT TTTGCATATG GAAATCACAG GCCCACCATT GGTACCATTT AATTCTACAA CAGTTATCAG CCTGCTAGTA GACAGTATAC GGGGCAAGAG ACTGGATGGC CGTGTCTTTA CCACACAAGA AAGTGTTGGG TCTAATGTTT AGATTGTTCC CTCAAAAGAG ATTCCTCAAA GCCAGTATGA

CTCTGCCTTC

GTAAGATGAT

ATTTATGTGA

ATTTAAGTTG

TGCCTGCTGG

ACAGCTGTTG

ATCCAAAACA

GCTCCTTATG

AATTCTTTAG

GCTAAAAGGA

AGAATCTATT

CCTGAAGTGT

GATGGTTCAC

CTGTGCCAAC

ACATCTGGTG

AAATATGGTG

ATGATGGTAA

CCACTTCAAT

AGAAGCTGAC

TTCCTGGGTC

CCAGGAATCT

TGTCATATAC

CT TT TCACTG

ATTGTTTGGT

TGT TGTGAGT CATC TATAAG AAGGAAAACA GGATGAATTT AGCTGAGAAA 91 9 9 9* 9 9 1251 1301 1351 1401 1451 .0 1501 1551 1601 1651 1701 1751 1801 1851 1901 1951 30 2001 2051 2101 2151 2201 2251 2301 2351 2401 2451 2501

GATCATGTTA

AGGAAAGTTT

ATCATOGCTA

TGTGAAACTG

TACAATCCAG

GCAGCCTTTA

AAAGATTGCT

AATCTTCCTA

TAGGTTCACT

AAGCCACTGC

ATTATTGAAA

AATTCCAGAT

TATGAGGTTT

C TTG TGTGCA TGGGATAT TG

ATAAAAGTTC

TACTATGAAA

AAAATGACTC

TCCTGCAATG

TTTCCTGTGG

CAATTGGTGC

AGCAAAGTAA

TTGTGTGATT

ATTTTATTAT

CTTAGAATCT

TATACTT

GCAAAG TTAC

ACCTATGATG

TGCTGAATTA

ATGGGTACT T

TCACTTGCGG

CTGTTCTGAT

ATTTGCAGAG

TTATCTGGCT

TGAC TC TCCA

OTOCATOCAG

ATATCTTGGG

TCGCTATGGT

ATGATGCAAA

GTCTATGCTG

GAGTAATTGG

CTATGAGAGG

T TT T TTCA.AT

CAGTGTACTG

TATGTGATTG

AACTAAAATG

AAAGCACTTT

ATTCCATCTA

TGATGGT TTT

ACACAATGTG

CCAACATGTG

TGTGAAAGCA

AAAAGCCAGT

TTAAGTLGGAA

ATCAAAATCT

TTCAGGTGCG

AGCAATCCAG

ACCTGAATTT

CTGAATGAAA

CTGCAATGAA

ATGTCAATAT

ACT TGCAGAT

GCAATTGAGG

TTCATCCCAT

TATGAATGCA

GATTAGGATC

TCCTTCCTC-A

GAXATTACTA

CTTTCCAGAG

AAGAAGTACA

GTCAGTCTCC

CTGTAAGAGG

CC TACACAG T

TGGAGAATAA

CCAAACCTCG

CATGAATGCC

CAATATCTCA

GGTCAACCAG

TATOATAGGA

ATCTGAGCCC

TTTTCCAGCC

AATCACTCTC

TTCTGTGGTG

TAAACATTGG

AATAACCTTC

ATGGAAGATG

CACT TCCAGA

CTAGATGGAC

TGTCATGGAT

TTAATGGAGA

AAGGAGAAAA ATGTCACTTT ACTTTGGAAG CCCCTGATGA ATTGTGCAGT GTTCAGAGAT ATGTGATAAA CCATCATACT GAACATGGTC AGAAGATGTG GGAAATCACA CGAAATTCAC ACAGAGCAAG CACATACTGT TACGGTTCTG GCCATCAATT TTCTGTTGCA AATTTTAATT TAACCTTTTC ATGGCCTATG ATATCGTGCA GTCACTCAGT GCTTATCCTT TAAACAGCAG GTTTCCTGGA TACTATCACC CAGTGATTAC AAGCTAATGT TGAGTGGAAA AATCTTAATG AAGATGGTGA AATAAAATGG CTTCATCTGT TAAGAAGTAT TATATCCATG GTAAGTTTAC 92 While the present invention has been described in terms of preferred embodiments, it is understood that variations and modifications will occur to those skilled in the art. Therefore, it is intended that the appended claims cover all such equivalent variations which come within the scope of the invention as claimed.

*9VV oo 93 SEQUENCE LISTING GENERAL INFORMATION: APPLICANT: CHANG, MING-SHI WELCHER, ANDREW A.

FLETCHER, FREDERICK A.

0 (ii) TITLE OF INVENTION: OB PROTEIN RECEPTOR AND RELATED COMPOSITIONS AND METHODS (iii) NUMBER OF SEQUENCES: 33 (iv) CORRESPONDENCE ADDRESS: ADDRESSEE: Amgen Inc.

STREET: 1840 Dehavilland Drive CITY: Thousand Oaks 0 STATE: California COUNTRY: USA ZIP: 91320 COMPUTER READABLE FORM: MEDIUM TYPE: Floppy disk COMPUTER: IBM PC compatible OPERATING SYSTEM: PC-DOS/MS-DOS SOFTWARE: PatentIn Release Version #1.30 0 (vi) CURRENT APPLICATION DATA: APPLICATION NUMBER: FILING DATE:

CLASSIFICATION:

(viii) ATTORNEY/AGENT

INFORMATION:

NAME: Pessin, Karol M.

S. REFERENCE/DOCKET NUMBER: A-382-A :0 INFORMATION FOR SEQ ID NO:1: SEQUENCE CHARACTERISTICS: LENGTH: 965 amino acids TYPE: amino acid STRANDEDNESS: single TOPOLOGY: linear (ii) MOLECULE TYPE: protein 94 (xi) SEQUENCE DESCRIPTION: SEQ ID NO:1: Met Ile Cys Gin Lys Phe Cys Val Vai Leu Leu His Trp Giu Phe Ile 1 5 10 Tyr Val Ile Thr Ala Phe Asn Leu Ser Tyr Pro Ile Thr Pro Trp Arg 25 0 Phe Lys Leu Ser Cys Met Pro Pro Asn Ser Thr Tyr Asp Tyr Phe Leu 40 Leu Pro Ala Gly Leu Ser Lys Asn Thr Ser Asn Ser Asn Gly His Tyr 55 Giu Thr Aia Val Giu Pro Lys Phe Asn Ser Ser Giy Thr His Phe Ser 70 75 Asn Leu Ser Lys Thr Thr Phe His Cys Cys Phe Arg Ser Giu Gin Asp 0 85 90 Arg Asn Cys Ser Leu Cys Ala Asp Asn Ile Giu Giy Lys Thr Phe Val 100 105 110 Ser Thr Val Asn Ser Leu Val Phe Gin Gin Ile Asp Ala Asn Trp Asn 115 120 125 .le Gin Cys Trp Leu Lys Giy Asp Leu Lys Leu Phe Ile Cys Tyr Val *5S130 135 140 **Giu Ser Leu Phe Lys Asn Leu Phe Arg Asn Tyr Asn Tyr Lys Vai His 145 150 155 160 Leu Leu Tyr Val Leu Pro Giu Vai Leu Giu Asp Ser Pro Leu Vai Pro 5165 170 175 Gin Lys Giy Ser Phe Gin Met Vai His Cys Asn Cys Ser Vai His Giu 5.*180 185 190 t0 Cys Cys Giu Cys- Leu Val Pro Vai Pro Thr Ala Lys Leu Asn Asp Thr 195 200 205 Leu Met Cys Leu Lys Ile Thr Ser Giy Gly Vai Ile Phe Gin Ser se.210 215 220 Pro Leu Met Ser Val Gin Pro Ile Asn Met Val Lys Pro Asp Pro Pro 225 230 235 240 Leu Gly Leu His Met Glu Ile Thr Asp Asp Gly Asn Leu Lys Ile Ser 0245 250 255 Trp Ser Ser Pro Pro Leu Vai Pro Phe Pro Leu Gin Tyr Gin Val Lys 260 265 270 95 Tyr Ser Giu 305 Asp Pro Ile T rp Ser 385 Pro Giu Asn T rp Arg 465 Pro Giu Ile Ser Ala 290 ValI T rp Pro Tyr Met 370 Asp Arg Cys Ile Ser 450 Tyr Ile Cys Arg Giu 275 Thr Gin Ser Lys Lys 355 Asn His Giy His Ser 435 Thr His Ser Ile Ile 515 Asn Se r Val1 Thr Ile 340 Lys Le u Val1 Lys His 420 Cys Ser Arg Giu Phe 500 As n Ser Leu Arg Pro 325 Leu Glu Ala Ser P he 405 Arg Giu Thr Ser Pro 485 Gin His Th r Le u Gly 310 Arg Th r Asn Glu Lys 390 Thr Tyr Thr Ile Se r 470 Lys Pro Ser Th r Val1 295 Lys Val1 Ser Lys Lys 375 Val1 Tyr Ala Asp Gin 455 Leu Asp Ile Leu Val1 280 Asp Arg Phe Val1 Ile 360 Ile Th r Asp Giu Gly 440 Ser Tyr Cys Phe Giy 520 Ser Le u Thr Gly 345 Val1 Pro Phe Ala Le u 425 Tyr Leu Cys Ty r Leu 505 Ser Ile Asp Thr 330 Se r Pro Gin Phe Val1 410 Tyr Leu Aia Ser Leu 490 Leu Leu Le u Gly 315 Gin As n Se r Ser Asn 395 Tyr Val1 Thr Giu Asp 475 Gin Ser Asp Pro 300 Pro Asp Val1 Lys Gin 380 Le u Cys Ile Lys Ser 460 Ile Se r Gly Ser Pro 540 Gly Gly Val1 Ser Giu 365 Tyr As n Cys Asp Met 445 Thr Pro Asp Tyr Pro 525 Ser Ile Ile Phe 350 Ile Asp Giu As n Val 430 Thr Leu Ser Giy Thr 510 Pro Ser T rp Tyr 335 His Val Val1 Thr Giu 415 As n Cys Gin Ile Phe 495 Met Thr Ty r Ser 320 Phe Cys T rp Vai Lys 400 His Ile Arg Leu His 480 Tyr Trp Cys Ile Arg Giu Ala Asp Lys Ile Vai 285 Val Leu Pro Asp Ser Val Vai Lys Pro Leu Pro 530 535 Ser Ser Vai Lys Ala 545 Pro )e Se r Val1 T rp 625 Arg Glu 2Leu *.*Gly T rp 705 Gly Lys 3 Cys Ty r T rp, 785 Phe Glu Glu ValI Gly Lys Gin 610 Se r Giy Lys Cys Th r 690 Thr Al a Val1 Val1 P he 770 Leu Ile Gly Ile P he Lys Se r 595 Vali As n Pro As n Se r 675 Trp Giu Se r Asn Ile 755 Ile Arg Pro Val Thr Ile Pro Giu 565 Giu Val 580 Vai Ser Arg Cys Pro Ala Giu Phe 645 Val Thr 660 Vai Gin Ser Giu Gin Ala Vai Aia 725 Ile Vai 740 Val Ser Ile Giu Ile Ser Ile Giu 805 Giy Lys 820 As n 550 Asn Gin Leu Lys Tyr 630 T rp Leu Arg Asp His 710 As n Gin T rp T rp Ser 790 Lys Pro Ile As n T rp Pro Arg Thr Arg Leu Tyr Val1 695 Thr Phe Ser Ile Lys 775 Ser Ty r Lys 96 Gly Leu Lys Val1 600 Leu Val1 Ile T rp Vali 680 Giy Val1 As n Leu Leu 760 As n Val1 Gin Ile Le u Gin Met 585 Pro Asp Vali Ile Lys 665 Ile Asn Thr Leu Se r 745 Ser Leu Lys Phe Ile 825 Leu P he 570 Ty r Asp Gly Met Asn 650 Pro As n His Vali Thr 730 Ala Pro As n Lys Ser 810 As n Lys 555 Gin Giu Leu Le u Asp 635 Gly Leu His Thr Leu 715 Phe Ty r Ser Giu Tyr 795 Leu Ser Ile Ile Vali Cys Giy 620 Ile Asp Met His Lys 700 Ala Ser Pro Asp Asp 780 Tyr Tyr Phe Se r Arg Tyr Al a 605 Tyr Lys Thr Lys Thr 685 P he Ile T rp Leu Tyr 765 Giy Ile Pro Thr T rp Tyr Asp 590 Val1 Trp ValI Met Asn 670 Se r Th r As n Pro Asn 750 Lys Giu His Ile Gin 830 Giu Gly 575 Ala Tyr Se r Pro Lys 655 Asp Cys P he Ser Met 735 Ser Leu Ile Asp Phe 815 Asp Lys 560 Leu Lys Ala As n Met 640 Lys Ser Asn Le u Ile 720 Se r Ser Met Lys His 800 Met Asp 97 Ile Giu Lys 835 Ile Ile Ser 850 His Gin Ser Asp Giy Leu Tyr Val Ile 845 Leu Vai Pro Vai Ile Ser His Ser Ser Ile Leu 855 Phe Leu Gly Thr Gin 865 Cys Arg Met Lys Lys Trp Giu Asp Val 875 Lys Asn Pro Lys Ser Trp Aia Gin 885 Ile Leu Asn Phe Arg Thr Asp Ile Leu 895 Ser Leu Ile Gin Ser Ile 915 Lys Lys Ile Thr Thr Asp Giu 905 Th r Asn Vai Pro Tyr Lys Ile Phe Arg Arg G ly 925 Gly Thr Ser Gin 910 Ala Asn Leu Leu Cys Phe Gin Leu Asn 930 Ara Thr Phe 935 Asn Leu Thr Tyr Asn Arg Cys Leu Gly Ser 945 Se r Lys 955 Arg Phe Giu Leu Asp Vai Leu 965 INFORMATION FOR SEQ ID NO:2: SEQUENCE CHARACTERISTICS: LENGTH: 3193 base pairs TYPE: nucieic acid STRANDEDNESS: singie TOPOLOGY: iinear (ii) MOLECULE TYPE: cDNA (Xi) SEQUENCE DESCRIPTION: SEQ ID NO:2: CCGCCGCCAT CTCTGCCTTC GGTCGAGTTG GACCCCCGGA TCAAGGTGTA CTTCTCTGAA GTAAGATGAT TTGTCAAAAA TTCTGTGTGG TTTTGTTACA TTGGGAATTT ATTTATGTGA TAACTGCGTT TAACTTGTCA TATCCAATTA CTCCTTGGAG ATTTAAGTTG TCTTGCATGC CACCAAATTC AACCTATGAC TACTTCCTTT TGCCTGCTGG ACTCTCAAAG AATACTTCAA ATTCGAATGG ACATTATGAG ACAGCTGTTG AACCTAAGTT TAATTCALAGT GGTACTCACT 98 a.

TTTCTAACTT

GCTCCTTATG

TTTTTCAACA

TATTCATCTG

TCCATCTTTT

0

GCAGTTTTCA

CTGTGCCAAC

GAGTAATTTT

CACCATTAGG

GCCCACCATT

0

CAGTTATCAG

TTCCTGGGTC

GGAGTGACTG

AAATTCTGAC

AGATTGTTCC

0

GCCAGTATGA

CCAA.ACCTCG

ATCATCGCTA

ATGGGTACTT

AAAGCACTTT

0

TTCATCCCAT

TTTTCCAGCC

TAGGTTCACT

CTCCATCCAG

AAAAGCCAGT

0

AAGAAGTACA

CAGTTCCAGA

ATCCAAAACA

TGCAGACAAC

AATAGATGCA

TTATGTGGAG

ATATGTTCTG

GATGGTTCAC

AGCCAA.ACTC

CCAGTCACCT

TTTGCATATG

GGTACCATTT

AGAAGCTGAC

TTCGTATGAG

GAGTACTCCT

AAGTGTTGGG

CTCAA.AAGAG

TGTTGTGAGT

AGGAAAGTTT

TGCTGAATTA

AACTAAAATG

GCAATTGAGG

ATCTGAGCCC

AATCTTCCTA

TGACTCTCCA

TGTGAAAGCA

CTTTCCAGAG

ATGGAAGATG

CTTGTGTGCA

ACTTTCCACT

ATTGAAGGAA

AACTGGAACA

TCATTATTTA

CCTGAAGTGT

TGCAAT'TGCA

AACGACACTC

CTAATGTCAG

GAAATCACAG

CCACTTCAAT

AAGATTGTCT

GTTCAGGTGA

CGTGTCTTTA

TCTAATGTTT

ATTGTTTGGT

GATCATGTTA

ACCTATGATG

TATGTGATTG

ACTTGCAGAT

TATCATAGGA

AAAGATTGCT

TTATCTGGCT

CCAACATGTG

GAAATTACTA

AATAACCTTC

TATGAGGTTT

GTCTATGCTG

GTTGCTTTCG

AGACATTTGT

TACAGTGCTG

AGAATCTATT

TAGAAGATTC

GTGTTCATGA

TCCTTATGTG

TTCAGCCCAT

ATGATGGTAA

ATCAAGTGAA

CAGCTACATC

GGGGCAAGAG

CCACACAAGA

CTTTTCACTG

GGATGAATT

GCAAAGTTAC

CAGTGTACTG

ATGTCAATAT

GGTCAACCAG

GCAGCCTTTA

ATTTGCAGAG

ACACAATGTG

TCCTTCCTGA

TAAACATTGG

AATTCCAGAT

ATGATGCAAA

TTCAGGTGCG

C-AGTGAGCAA

TTCALACAGTA

GCTAAAAGGA

CAGGAATTAT

ACCTCTGGTT

ATGTTGTGAA

TTTGAAAATC

AAATATGGTG

TTTAAAGATT

ATATTCAGAG

CCTGCTAGTA

ACTGGATGGC

TGTCATATAC

CATCTATAAG

AGCTGAGAAA

TTTTTTCAAT

CTGCAATGAA

CAATATCTCA

TACAATCCAG

CTGTTCTGAT

TGATGGTTTT

GATTAGGATC

TTCTGTGGTG

ATTATTGAAA

TCGCTATGGT

ATCAAAATCT

CTGTAAGAGG

GATAGAAACT

AATTCTTTAG

GACTTAAAAT

AACTATAAGG

CCCCAAAAAG

TGTCTTGTGC

ACATCTGGTG

AAGCCTGATC

TCTTGGTCCA

AATTCTACAA

GACAGTATAC

CCAGGAATCT

TTTCCACCTA

AAGGAAAACA

ATTCCTCAAA

CTGAATGAAA

CATGAATGCC

TGTGAAACTG

TCACTTGCGG

ATTCCATCTA

TATGAATGCA

AATCACTCTC

AAGCCACTGC

ATATCTTGGG

TTAAGTGGAA

GTCAGTCTCC

CTAGATGGAC

360 420 480 540 600 660 720 780 840 900 960 1020 1080 1140 1200 1260 1320 1380 1440 1500 1560 1620 1680 1740 1800 1860 1920 99

TGGGATATTG

CTATGAGAGG

ATGTCACTTT

ATGTGATAAA

.0 CGAAATTCAC

CAATTGGTGC

ATATCGTGCA

TACTATCACC

AAGATGGTGA

?0 ATCATTTTAT

TGGGAAAACC

ATGCAGGTTT

CATTATTAAT

AGAATTGTTC

TAATCATGAT

AGAAGATTTT

GAGAGTTAAC

GTTCAAAATG

TTTAAAAGTA

4 0 TTAAGAAAAT

TAGACAATTT

GAGTAATTGG

ACCTGAATTT

ACTTTGGAAG

CCATCATACT

TTTCCTGTGG

TTCTGTTGCA

GTCACTCAGT

CAGTGATTAC

AATAAAATGG

CCCCATTGAG

AAAGATAATT

ATATGTAATT

ATCACACCAA

CTGGGCACAA

CACTACAGAT

TACATTTTGA

ATATGGTGGA

TAGATTTGAG

GTATTCATGA

TATGGCTGTT

TGTAATTAGG

AGCAATCCAG

TGGAGAATA

CCCCTGATGA

TCCTGCAATG

ACAGAGCAAG

AATTTTAATT

GCTTATCCTT

AAGCTAATGT

CTTAGAATCT

AAGTACCAGT

AATAGTTTCA

GTGCCAGTAA

AGAATGAAAA

GGACTTAATT

GAACCCAATG

AGAAGGGGAG

TTATGTTGAT

TCCAGTTTGG

TTTCTGGCTT

GCTGTCATTA

TGAACTCTAA

CCTACACAGT

TTAATGGAGA

AAAATGACTC

GAACATGGTC

CACATACTGT

TAACCTTTTC

TAAACAGCAG

ATTTTATTAT

CTTCATCTGT

TCAGTCTTTA

CTCAAGATGA

TTATTTCCTC

AGCTATTTTG

TTCAGAAGAG

TGCCAACTTC

CAAATCTAAA

TTAGAACTTA

ATGTGTGATT

TTGATTTGCC

CATATCTATT

AACTGCAACA

TGTCATGGAT

TACTATGAAA

ATTGTGCAGT

AGAAGATGTG

TACGGTTCTG

ATGGCCTATG

TTGTGTGATT

TGAGTGGAAA

TAAGAAGTAT

CCCAATATTT

TATTGAAAAA

TTCCATCTTA

GGAAGATGTT

AACGGACATT

CCAACAGTCT

AAAAATTCAG

AAATAGATGT

AATTTTCAAA

ATATTCCTGG

AAATGTCATC

TCTGACAAAT

ATAAAAGTTC

AAGGAGAAAA

GTTCAGAGAT

GGAAATCACA

GCCATCAATT

AGCAAAGTAA

GTTTCCTGGA

AATCTTAATG

TATATCCATG

ATGGAAGGAG

CACCAGAGTG

TTGCTTGGAA

CCGAACCCCA

CTTTGAAGTC

ATAGAGTATT

TTGAACTTCT

GTAAATTTGG

TCATCTAAAG

TCATAAAACA

AAATATGTAG

TGCTTTAAAA

1980 2040 2100 2160 2220 2280 2340 2400 2460 2520 2580 2640 2700 2760 2820 2880 2940 3000 3060 3120 3180 3193 ATACAATGAT TAT INFORMATION FOR SEQ ID NO:3: SEQUENCE CHARACTERISTICS: LENGTH: 995 amino acids TYPE: amino acid STRANDEDNESS: single TOPOLOGY: linear 100- (ii) MOLECULE TYPE: protein (xi) SEQUENCE DESCRIPTION: SEQ ID NO:3: Met Ile Cys Gin Lys Phe Cys Val Vai Leu Leu His Trp Giu Phe Ile 1 5 10 .0 Tyr Vai Ile Thr Ala Phe Asn Leu Ser Tyr Pro Ile Thr Pro Trp Arg 25 Phe Lys Leu Ser Cys Met Pro Pro Asn Ser Thr Tyr Asp Tyr Phe Leu L535 40 Leu Pro Ala Giy Leu Ser Lys Asn Thr Ser Asn Ser Asn Gly His Tyr 55 0 Giu Thr Ala Val Giu Pro Lys Phe Asn Ser Ser Giy Thr His Phe Ser 70 75 Asn Leu Ser Lys Thr Thr Phe His Cys Cys Phe Arg Ser Giu Gin Asp 90 Arg Asn Cys Ser Leu Cys Ala Asp Asn Ile Giu Giy Lys Thr Phe Val *100 105 110 Ser Thr Val Asn Ser Leu Vai Phe Gin Gin Ile Asp Ala Asn Trp Asn 115 120 125 Ile Gin Cys Trp Leu Lys Gly Asp Leu Lys Leu Phe Ile Cys Tyr Val 130 135 140 Giu Ser Leu Phe Lys Asn Leu Phe Arg Asn Tyr Asn Tyr Lys Val His *145 150 155 160 Leu Leu Tyr Vai Leu Pro Glu Val Leu Giu Asp Ser Pro Leu Vai Pro 165 170 175 Gin Lys Gly Ser Phe Gin Met Val His Cys Asn Cys Ser Val His Giu *180 185 190 Cys Cys Giu Cys Leu Vai Pro Vai Pro Thr Ala Lys Leu Asn Asp Thr 195 200 205 Leu Leu Met Cys Leu Lys Ile Thr Ser Giy Gly Vai Ile Phe Gin Ser 210 215 220 Pro Leu Met Ser Vai Gin Pro Ile Asn Met Val Lys Pro Asp Pro Pro 225 230 235 240 101 Leu Gly Leu His Met Giu Ile Thr Asp Asp Gly Asn Leu Lys Ile Ser 245 250 255 Trp Ser Ser Pro Pro Leu Val Pro Phe Pro Leu Gin Tyr Gin Vai Lys 260 265 270 Tyr Ser Giu Asn Ser Thr Thr Vai Ile Arg Giu Ala Asp Lys Ile Val 275 280 285 LO Ser Ala Thr Ser Leu Leu Val Asp Ser Ile Leu Pro Gly Ser Ser Tyr 290 295 300 Giu Val Gin Vai Arg Giy Lys Arg Leu Asp Gly Pro Giy Ile Trp Ser 305 310 315 320 Asp Trp Ser Thr Pro Arg Vai Phe Thr Thr Gin Asp Vai Ile Tyr Phe 325 330 335 Pro Pro Lys Ile Leu Thr Ser Vai Gly Ser Asn Vai Ser Phe His Cys 340 345 350 Ile Tyr Lys Lys Giu Asn Lys Ile Vai Pro Ser Lys Giu Ile Vai Trp 355 360 365 25 Trp Met Asn Leu Ala Giu Lys Ile Pro Gin Ser Gin Tyr Asp Vai Val .370 375 380 Ser Asp His Val Ser Lys Val Thr Phe Phe Asn Leu Asn Giu Thr Lys **385 390 395 400 Pro Arg Gly Lys Phe Thr Tyr Asp Ala Val Tyr Cys Cys Asn Giu His *405 410 415 Giu Cys His His Arg Tyr Ala Giu Leu Tyr Val Ile Asp Val Asn Ile 420 425 430 Asn Ile Ser Cys Giu Thr Asp Gly Tyr Leu Thr Lys Met Thr Cys Arg 435 440 445 40 Trp Ser Thr Ser Thr Ile Gin Ser Leu Ala Giu Ser Thr Leu Gin Leu 450 455 460 Arg Tyr His Arg Ser Ser Leu Tyr Cys Ser Asp Ile Pro Ser Ile His 465 470 475 480 Pro Ile Ser Giu Pro Lys Asp Cys Tyr Leu Gin Ser Asp Gly Phe Tyr 485 49049 Giu Cys Ile Phe Gin Pro Ile Phe Leu Leu Ser Giy Tyr Thr Met Trp 500 505 510 Ile Arg Ile Asn His Ser Leu Gly Ser Leu Asp Ser Pro Pro Thr Cys 515 52052 525 102 Val Ala 545 Pro Ser Ser Val Trp 625 Arg Glu Leu Gly Trp 705 Gly Lys Cys Tyr Leu 530 Glu Val Gly Lys Gin 610 Ser Gly Lys Cys Thr 690 Thr Ala Val Val Phe Pro Ile Phe Lys Ser 595 Val Asn Pro Asn Ser 675 Trp Glu Ser Asn Ile 755 Ile Asp Thr Pro Glu 580 Val Arg Pro Glu Val 660 Val Ser Gin Val Ile 740 Val Ile Ser Ile Glu 565 Val Ser Cys Ala Phe 645 Thr Gin Glu Ala Ala 725 Val Ser Glu Val Asn 550 Asn Gin Leu Lys Tyr 630 Trp Leu Arg Asp His 710 Asn Gin Trp Trp Val 535 Ile Asn Trp Pro Arg 615 Thr Arg Leu Tyr Val 695 Thr Phe Ser Ile Lys 775 Lys Gly Leu Lys Val 600 Leu Val Ile Trp Val 680 Gly Val Asn Leu Leu 760 Asn Leu Gin Met 585 Pro Asp Val Ile Lys 665 Ile Asn Thr Leu Ser 745 Ser Leu Leu Phe 570 Tyr Asp Gly Met Asn 650 Pro Asn His Val Thr 730 Ala Pro Asn Lys 555 Gin Glu Leu Leu Asp 635 Gly Leu His Thr Leu 715 Phe Tyr Ser Glu Ile Ile Val Cys Gly 620 Ile Asp Met His Lys 700 Ala Ser Pro Asp Asp 780 Ser Arg Tyr Ala 605 Tyr Lys Thr Lys Thr 685 Phe Ile Trp Leu Tyr 765 Gly Trp Tyr Asp 590 Val Trp Val Met Asn 670 Ser Thr Asn Pro Asn 750 Lys Glu Glu Gly 575 Ala Tyr Ser Pro Lys 655 Asp Cys Phe Ser Met 735 Ser Leu Ile Lys 560 Leu Lys Ala Asn Met 640 Lys Ser Asn Leu Ile 720 Ser Ser Met Lys Pro Leu Pro Pro Ser Ser Val Lys 540 770 Trp 785 Leu Arg Ile Ser Ser Ser Val Lys Lys Tyr Tyr Ile His Asp His 790 795 800 103 Phe Ile Pro Ile Glu Lys Tyr Gin Phe Ser Leu Tyr Pro Ile Phe Met 805 810 815 Glu Gly Val Gly Lys Pro Lys Ile Ile Asn Ser Phe Thr Gin Asp Asp 820 825 830 Ile Glu Lys His Gin Ser Asp Ala Gly Leu Tyr Val Ile Val Pro Val 835 840 845 LO Ile Ile Ser Ser Ser Ile Leu Leu Leu Gly Thr Leu Leu Ile Ser His 850 855 860 Gin Arg Met Lys Lys Leu Phe Trp Glu Asp Val Pro Asn Pro Lys Asn 865 870 875 880 Cys Ser Trp Ala Gin Gly Leu Asn Phe Gin Lys Lys Arg Leu Ser Ile 885 890 895 Phe Leu Ser Ser Ile Gin His Gin His Val Val Leu Phe Phe Trp Ser 900 905 910 Leu Lys Gin Phe Gin Lys Ile Ser Val Leu Ile His His Gly Lys Ile 915 920 925 25 Lys Met Arg Cys Gin Gin Leu Trp Ser Leu Tyr Phe Gin Gin Gin Ile 930 935 940 Leu Lys Arg Val Leu Phe Val Leu Val Thr Ser Ser Thr Val Leu Thr 945 950 955 960 Ser Leu Arg Leu Arg Val Leu Arg Pro Met Arg Thr Lys Ala Arg Asp 965 970 975 Asn Pro Leu Leu Asn Thr Pro Arg Ser Ala Thr Leu Asn Gln Val Lys 980 985 990 Leu Val Lys 995 40 INFORMATION FOR SEQ ID NO:4: SEQUENCE CHARACTERISTICS: LENGTH: 3063 base pairs TYPE: nucleic acid STRANDEDNESS: single TOPOLOGY: linear (ii) MOLECULE TYPE: cDNA 104 (xi) SEQUENCE DESCRIPTION: SEQ ID NO:4: CCGCCGCCAT CTCTGCCTTC GGTCGAGTTG GACCCCCGGA TCAAGGTGTA CTTCTCTGAA V 0 0 0 3 GTAAGATGAT TTGTCAAAAA TAACTGCGTT TAACTTGTCA CACCAAATTC AACCTATGAC ATTCGAATGG ACATTATGAG TTTCTAACTT ATCCAAAACA GCTCCTTATG TGCAGACAAC TTTTTCAACA AATAGATGCA TATTCATCTG TTATGTGGAG 0 TCCATCTTTT ATATGTTCTG GCAGTTTTCA GATGGTTCAC CTGTGCCA-AC AGCCAAACTC GAGTAATTTT CCAGTCACCT CACCATTAGG TTTGCATATG 0 GCCCACCATT GGTACCATTT CAGTTATCAG AGAAGCTGAC TTCCTGGGTC TTCGTATGAG GGAGTGACTG GAGTACTCCT AAATTCTGAC AAGTGTTGGG 0 AGATTGTTCC CTCAAAAGAG GCCAGTATGA TGTTGTGAGT CCAAACCTCG AGGAAAGTTT ATCATCGCTA TGCTGAATTA ATGGGTACTT AACTAAAATG AAAGCACTTT GCAATTGAGG TTCATCCCAT ATCTGAGCCC

TTCTGTGTGG

TATCCAATTA

TACTTCCTTT

ACAGCTGTTG

ACTTTCCACT

ATTGAAGGAA

AACTGGAACA

TCATTATTTA

CCTGAAGTGT

TGCAATTGCA

AACGACACTC

CTAATGTCAG

GAAATCACAG

CCACTTCAAT

AAGATTGTCT

GTTCAGGTGA

CGTGTCTTTA

TCTAATGTTT

ATTGTTTGGT

GATCATGTTA

ACCTATGATG

TATGTGATTG

ACTTGCAGAT

TATCATAGGA

AAAGATTGCT

TTTTGTTACA

CTCCTTGGAG

TGCCTGCTGG

AACCTAAGTT

GTTGCTTTCG

AGACATTTGT

TACAGTGCTG

AGAATCTATT

TAGAAGATTC

GTGTTCATGA

TCCTTATGTG

TTCAGCCCAT

ATGATGGTAA

ATCAAGTGAA

CAGCTACATC

GGGGCA.AGAG

CCACACAAGA

CTTTTCACTG

GGATGAATTT

GCAAAGTTAC

CAGTGTACTG

ATGTCAATAT

GGTCAACCAG

GCAGCCTTTA

ATTTGCAGAG

TTGGGAATTT

A.TTTAAGTTG

ACTCTCAAAG

TAATTCAAGT

GAGTGAGCAA

TTCAACAGTA

GCTAAAAGGA

CAGGAATTAT

ACCTCTGGTT

ATGTTGTGAA

TTTGAAAATC

AAATATGGTG

TTTAAAGATT

ATATTCAGAG

CCTGCTAGTA

ACTGGATGGC

TGTCATATAC

CATCTATAAG

AGCTGAGAAA

TTTTTTCAAT

CTGCAATGAA

CAATATCTCA

TACAATCCAG

CTGTTCTGAT

TGATGGTTTT

ATTTATGTGA

TCTTGCATGC

AATACTTCAA

GGTACTCACT

GATAGAAACT

AATTCTTTAG

GACTTAAAAT

AACTATAAGG

CCCCAAAAAG

TGTCTTGTGC

ACATCTGGTG

AAGCCTGATC

TCTTGGTCCA

AATTCTACAA

GACAGTATAC

CCAGGAATCT

TTTCCACCTA

A AGGAAAACA

ATTCCTCAAA

CTGAATGAAA

CATGAATGCC

TGTGAAACTG

TCACTTGCGG

ATTCCATCTA

TATGAATGCA

120 180 240 300 360 420 480 540 600 660 720 780 840 900 960 1020 1080 1140 1200 1260 1320 1380 1440 1500 1560 S. 55 105

TTTTCCAGCC

TAGGTTCACT

CTCCATCCAG

AAAAGCCAGT

0 AAGAAGTACA

CAGTTCCAGA

TGGGATATTG

CTATGAGAGG

ATGTCACTTT

ATGTGATAAA

CGAAATTCAC

CAATTGGTGC

ATATCGTGCA

TACTATCACC

AAGATGGTGA

ATCATTTTAT

TGGGAAAACC

ATGCAGGTTT

CATTATTAAT

10 AGAATTGTTC

CAAGCATACA

AGATATCAGT

CTCTCTACTT

CAGTGTTAAC

0o ACA.ACCCTTT

AGA

AATCTTCCTA

TGACTCTCCA

TGTGAA.AGCA

CTTTCCAGAG

ATGGAAGATG

CTTGTGTGCA

GAGTAATTGG

ACCTGAATTT

ACTTTGGAAG

CCATCATACT

TTTCCTGTGG

TTCTGT-TGCA

GTCACTCAGT

CAGTGATTAC

AATAAAATGG

CCCCATTGAG

AAAGATAATT

ATATGTAATT

ATCACACCAA

CTGGGCACAA.

GCATCAGTGA

GTTGATACAT

TCAACAACAG

TTCTCTGAGG

GTTAAATACG

TTATCTGGCT

CCAACATGTG

GAAATTACTA

AATAACCTTC

TATGAGGTTT

GTCTATGCTG

AGCAATCCAG

TGGAGAATAA

CCCCTGATGA

TCCTGCAATG

ACAGAGCAAG

ALATTTTAATT

GCTTATCCTT

AAGCTAATGT

CTTAGAATCT

AAGTACCAGT

AATAGTTTCA

GTGCCAGTAA

AGAATGAAAA

GGACTTAATT

CATGTGGTCC

CATGGAAAAA

ATCTTGAAAA

CTGAGGGTAC

CCACGCTGAT

ACACAATGTG

TCCTTCCTGA

TAAACATTGG

AATTCCAGAT

ATGATGCAAA

TTCAGGTGCG

CCTACACAGT

TTAATGGAGA

AAAATGACTC

GAACATGGTC

CACATACTGT

TAACCTTTTC

TAAACAGCAG

ATTTTATTAT

CTTCATCTGT

TCAGTCTTTA

CTCAAGATGA

TTATTTCCTC

AGCTATTTTG

TTCAGAAGAA

TCTTCTTTTG

TAAAGATGAG

GGGTTCTGTT

TGAGGTAACC

CAGCAACTCT

GATTAGGATC

TTCTGTGGTG

ATTATTGAAA

TCGCTATGGT

ATCAAAATCT

CTGTA.AGAGG

TGTCATGGAT

TACTATGAAA

ATTGTGCAGT

AGAAGATGTG

TACGGTTCTG

ATGGCCTATG

TTGTGTGATT

TGAGTGGAAA

TAAGAAG TAT

CCCA.ATATTT

TATTGAAAAA

TTCCATCTTA

GGAAGATGTT

ACGTTTGAGC

GAGCCTGAAA

ATGATGCCAA

TGTTTTAGTG

TATGAGGACG

AAACCAAGTG

AATCACTCTC

AAGCCACTGC

ATATCTTGGG

TTAAGTGGAA

GTCAGTCTCC

CTAGATGGAC

ATAAAAGTTC

AAGGAGAAAA

GTTCAGAGAT

GGAAATCACA

GCCATCAATT

AGCAAAGTAA

GTTTCCTGGA

AATCTTAATG

TATATCCATG

ATGGAAGGAG

CACCAGAGTG

TTGCTTGGAA

CCGAACCCCA

ATCTTTTTAT

CAATTTCAGA

CAACTGTGGT

ACCAGTTCAA

AAAGCCAGAG

AAACTGGTGA

1620 1680 1740 1800 1860 1920 1980 2040 2100 2160 2220 2280 2340 2400 2460 2520 2580 2640 2700 2760 2820 2880 2940 3000 3060 3063 106 INFORMATION FOR SEQ ID SEQUENCE CHARACTERISTICS: LENGTH: 969 amino acids TYPE: amino acid STRANDEDNESS: single TOPOLOGY: linear (ii) MOLECULE TYPE: protein (xi) SEQUENCE DESCRIPTION: SEQ ID Met Ile Cys Gin Lys Phe Cys Val Val Leu Leu His Trp Glu Phe Ile 0 0000 0 0000 0000 00 00 0 00 0 0 *0 0900..

0 0 000000 0 000.

0 0 000e 90 0 00 0* 0000 0 0000 00 0000 0 0000 0 9 0000 1 Tyr P he Leu Glu 65 Asn Arg Ser Ile Glu 145 Leu Gin V.ali Lys Pro 50 Thr Leu As n Thr Gin 130 Ser Leu Lys Ile Leu 35 Ala Ala Ser Cys Val 115 Cys Leu Tyr Gly Thr Ser G1y Val1 Lys Ser 100 Asn T rp Phe *Val *Ser 180 Al a Cys Leu Glu Thr 85 Leu Ser Leu Lys Leu 165 P he Phe Met Se r Pro 70 Thr Cys Leu Lys As n 150 Pro Gin .s n Pro Ly s 55 Lys Phe Al a Val1 Gly 135 Leu Giu Met Leu Pro 40 Asn P he His Asp P he 120 Asp Phe Val1 Val Ser 25 Asn Thr As n Cys As n 105 Gin Leu Arg Leu His 185 Ty r Se r Se r Se r Cys 90 Ile Gin Lys As n Glu 170 Cys Pro Thr As n Se r 75 Phe Glu Ile Leu Ty r 155 Asp As n Ile Tyr Se r Gly Arg Gly Asp Phe 140 Asn Se r Cys Thr Asp As n Thr Ser Lys Ala 125 Ile Tyr Pro Se r Pro Tyr Giy His Glu Thr 110 As n Cys Lys Leu Val1 Trp, P he His Phe Gin Phe Trp Tyr Val1 Vai 175 His Arg Leu Tyr Ser Asp Val1 As n Val1 His 160 Pro Glu 190 Cys Cys Glu Cys Leu Val Pro Val Pro Thr Ala Lys Leu Asn Asp Thr 195 200 205 107 Leu Leu Met Cys Leu Lys Ile Thr Ser Giy Giy Val Ile Phe Gin Ser 210 215 220 Leu Met Ser Val Gin Pro Ile Asn Met Vai Lys Pro Asp Pro Pro 225 230 235 240 Leu Giy Leu His Met Giu Ile Thr Asp Asp Gly Asn Leu Lys Ile Ser 245 250 255 Trp Ser Ser Pro Pro Leu Vai Pro Phe Pro Leu Gin Tyr Gin Val Lys 260 265 270 Tyr Ser Giu Asn Ser Thr Thr Vai Ile Arg Giu Aia Asp Lys Ile Val 5275 280 285 Ser Aia Thr Ser Leu Leu Vai Asp Ser Ile Leu Pro Gly Ser *Ser Tyr 290 295 300 0 Giu Val Gin Vai Arg Giy Lys Arg Leu Asp Giy Pro Giy Ile Trp Ser 305 310 315 320 Asp Trp, Ser Thr Pro Arg Vai Phe Thr Thr Gin Asp Vai Ile Tyr Phe 325 330 335 Pro Pro Lys Ile Leu Thr Ser Vai Giy Ser Asn Vai Ser Phe His Cys 340 345 350 Sle Tyr Lys Lys Giu Asn Lys Ile Val Pro Ser Lys Giu Ile Vai Trp 0 355 360 365 *Trp Met Asn Leu Ala Giu Lys Ile Pro Gin Ser Gin Tyr Asp Val Val 370 375 380 15 Ser Asp His Vai Ser Lys Vai Thr Phe Phe Asn Leu Asn Glu Thr Lys 385 390 395 400 *:Pro Arg Gly Lys Phe Thr Tyr Asp Ala Val Tyr Cys Cys Asn Glu His Giu Cys His His Arg Tyr Ala Giu Leu Tyr Val Ile Asp Val Asn Ile 420 425 430 Asn Ile Ser Cys Glu Thr Asp Gly Tyr Leu Thr Lys met Thr Cys Arg 15435 440 445 Trp, Ser Thr Ser Thr Ile Gin Ser Leu Ala Glu Ser Thr Leu Gin Leu 450 455 460 Arg Tyr His Arg Ser Ser Leu Tyr Cys Ser Asp Ile Pro Ser Ile His 465 470 475 480 108 Pro Ile Ser Giu Pro Lys Asp Cys 1 Glu Ile Val Ala 545 Pro Ser Ser Val Trp 625 Arg Glu Leu Gly Trp 705 Gly Lys -ys Arg Leu 530 lu Va1 Gly Lys Gin 610 Ser Gly Lys Cys Th-r 690 Thr Ala Val Ile Ile 515 Pro Ile Phe Lys Ser 595 Val Asn Pro Asn Ser 675 Trp Glu Ser Asn Phe 500 Asn I Asp Thr Pro Glu 580 Va1 Arg Pro Glu Va1 660 Val Ser Gin Val Ile 740 3In -is Ser Ile Glu 565 Val Ser Cys Ala Phe 645 Thr Gin Glu Ala Ala 725 Val Pro Ser Val Asn 550 Asn Gin Leu Lys Tyr 630 Trp Leu Arg Asp His 710 Asn Gin Lie I Leu Val 535 Ile Asn Trp Pro Arg 615 Thr Arg Leu Tyr Vai 695 Thr Phe Ser ?he 520 Lys Gly Leu Lys Val 600 Leu Va1 Ile Trp Val 680 Gly Val Asr Let Tyr I 4 Leu I 505 Ser I Pro I Leu Gin Met 585 Pro Asp Val' Ile Lys 665 Ile Asn Thr Leu i Ser 745 ,eu 190 .eu ,eu Leu Leu ?he 570 Tyr Asp Gly Met Asn 650 Pro Asn His Val Thi 730 Alz Gin S Ser C Asp Pro Lys 555 Gin Glu Leu Leu Asp 635 Gly Leu His Thr Leu 715 Phe Tyr er ;ly 3er ?ro 540 Ile Ile Vai Cys Gly 620 Ile Asp Met His Lys 700 Ala Sex Pro Aso Tyr Pro 525 Ser Ser Arg Tyr Ala 605 Tyr Lys Thr Lys Thr 685 Phe Ile TrF Let Gly I Thr 510 Pro Ser Trp Tyr Asp 590 Val Trp Val Met Asn 670 Ser Thr Asn Pro a Asn 750 ?he 195 det rhr Val Slu Gly 575 Ala Tyr Ser Pro Lys 655 Asp Cys Phe Ser Met 735 Sei Tyr Trp Cys Lys Lys 560 Leu Lys Ala Asn Met 640 Lys Ser Asn Leu Ile 720 Ser Ser Cys Val Ile 755 Val Ser Trp Ile Ser Pro Ser Asp Tyr 765 Lys Leu Met 109 Tyr Phe Ile Ile Giu Trp Lys Asn Leu Asn Giu Asp Gly Giu Ile Lys 770 775 780 D Trp Leu Arg Ile Ser Ser Ser Vai Lys Lys Tyr Tyr Ile His Asp His 785 790 795 800 Phe Ile Pro Ile Giu Lys Tyr Gin Phe Ser Leu Tyr Pro Ile Phe Met 805 810 815 Giu Gly Vai Gly Lys Pro Lys Ile Ile Asn Ser Phe Thr Gin Asp Asp 820 825 830 Ile Giu Lys His Gin Ser Asp Aia Giy Leu Tyr Vai Ile Vai Pro Val 835 840 845 Ile Ile Ser Ser Ser Ile Leu Leu Leu Giy Thr Leu Leu Ile Ser His 850 855 860 0 Gin Arg Met Lys Lys Leu Phe Trp Giu Asp Val Pro Asn Pro Lys Asn 865 870 875 880 Cys Ser Trp Ala Gin Giy Leu Asn Phe Gin Lys Met Leu Giu Giy Ser 885 890 895 Met Phe Val Lys Ser His His His Ser Leu Ile Ser Ser Thr Gin Giy 900 905 910 *His Lys His Cys Giy Arg Pro Gin Giy Pro Leu His Arg Lys Thr Arg X00915 920 925 *.Asp Leu Cys Ser Leu Vai Tyr Leu Leu Thr Leu Pro Pro Leu Leu Ser 930 935 940 5 Tyr Asp Pro Ala Lys Ser Pro Ser Val Arg Asn Thr Gin Glu Ser Ile 945 950 955 960 *:Lys Lys Lys Lys Lys Lys Leu Giu Giy INFOMTION FOR SEQ ID NO:6: SEQUENCE CHARACTERISTICS: LENGTH: 969 amino acids TYPE: amino acid STRANDEDNESS: single TOPOLOGY: linear (ii) MOLECULE TYPE: protein J110 (xi) SEQUENCE DESCRIPTION: SEQ ID NO:6: Met Ile Cys Gin Lys Phe Cys Val Vai Leu Leu His Trp Giu Phe Ile 1 5 10 Tyr Vai Ile Thr Ala Phe Asn Leu Ser Tyr Pro Ile Thr Pro Trp Arg 25 Phe Lys Leu Ser Cys Met Pro Pro Asn Ser Thr Tyr Asp Tyr Phe Leu 335 40 Leu Pro Ala Gly Leu Ser Lys Asn Thr Ser Asn Ser Asn Giy His Tyr 55 Thr Ala Vai Giu Pro Lys Phe Asn Ser Ser Giy Thr His Phe Ser 70 75 Asn Leu Ser Lys Thr Thr Phe His Cys Cys Phe Arg Ser Giu Gin Asp 90 Arg Asn Cys Ser Leu Cys Aia Asp Asn Ile Giu Gly Lys Thr Phe Val 100 105 110 Ser Thr Vai Asn Ser Leu Vai Phe Gin Gin Ile Asp Ala Asn Trp Asn 5115 120 125 Sle Gin Cys Trp Leu Lys Gly Asp Leu Lys Leu Phe Ile Cys Tyr Val 130 135 140 0Giu Ser Leu Phe Lys Asn Leu Phe Arg Asn Tyr Asn Tyr Lys Val His *145 150 155 160 Leu Leu Tyr Val Leu Pro Giu Val Leu Glu Asp Ser Pro Leu Val Pro 165 170 175 Gin Lys Gly Ser Phe Gin Met Val His Cys Asn Cys Ser Vai His Giu 180 185 190 :Cys Cys Giu Cys Leu Vai Pro Val Pro Thr Ala Lys Leu Asn Asp Thr 0195 200 205 Leu Leu Met Cys Leu Lys Ile Thr Ser Gly Gly Vai Ile Phe Gin Ser 210 215 220 5Pro Leu Met Ser Val Gin Pro Ile Asn Met Val Lys Pro Asp Pro Pro 225 230 235 240 Leu Giy Leu His Met Giu Ile Thr Asp Asp Gly Asn Leu Lys Ile Ser 245 250 255 Trp Ser Ser Pro Pro Leu Val Pro Phe Pro Leu Gin Tyr Gin Vai Lys 260 265 270 ill Tyr Ser GJlu Asn Ser Thr Thr Val Ile Arg Giu Ala Asp Lys Ile Vai 275 280 285 Ser Ala Thr Ser Leu Leu Val Asp Ser Ile Leu Pro Gly Ser Ser Tyr 290 295 300 Giu Vai Gin Val Arg Giy Lys Arg Leu Asp Gly Pro Gly Ile Trp Ser 305 310 315 320 Asp Trp Ser Thr Pro Arg Val Phe Thr Thr Gin Asp Val Ile Tyr Phe 325 330 335 Pro Pro Lys Ile Leu Thr Ser Val Giy Ser Asn Val. Ser Phe H-is Cys 340 345 350 Ile Tyr Lys Lys Giu Asn Lys Ile Val Pro Ser Lys Giu Ile Vai Trp 355 360 365 Trp Met Asn Leu Aia Giu Lys Ile Pro Gin Ser Gin Tyr Asp Val Val 370 375 380 Ser Asp His Val Ser Lys Val Thr Phe Phe Asn Leu Asn Giu Thr Lys 385 390 395 400 Pro Arg Giy Lys Phe Thr Tyr Asp Ala Vai Tyr Cys Cys Asn Giu His 405 410 415 Giu Cys His His Arg Tyr Ala Giu Leu.Tyr Val Ile Asp Vai Asn Ile *420 425 430 Asn Ile Ser Cys Giu Thr Asp Gly Tyr Leu Thr Lys Met Thr Cys Arg *435 440 445 Trp Ser Thr Ser Thr Ile Gin Ser Leu Ala Giu Ser Thr Leu Gin Leu 35 450 455 460 Arg Tyr His Arg Ser Ser Leu Tyr Cys Ser Asp Ile Pro Ser Ile His 465 470 475 480 40 Pro Ile Ser Giu Pro Lys Asp Cys Tyr Leu Gin Ser Asp Gly Phe Tyr 485 490 495 Giu Cys Ile Phe Gin Pro Ilie Phe Leu Leu Ser Giy Tyr Thr Met Trp 50*1 Ile Arg Ile Asn His Ser Leu Giy Ser Leu Asp Ser Pro Pro Thr Cys 515 520 525 Val Leu Pro Asp Ser Vai Vai Lys Pro Leu Pro Pro Ser Ser Vai Lys 530 535 540 Ala Giu Ile Thr Ile Asn Ile Gly Leu Leu Lys Ile Ser Trp Giu Lys 545 550 55560 112 Pro Ser Ser Va1 Trp 625 Arg Glu Leu Gly Trp 705 Gly Lys Cys Tyi Trj 78~ Phe Val P Gly L Lys S Gin 1 610 Ser Gly I Lys Cys Thr 690 Thr Ala Val Val Phe 770 Leu B Ile he lys jer la1 ksn ?ro Asn Ser 675 Trp Glu Set Asn Ile 755 Ile Arg Prc Pro G Glu 580 Val Arg Pro Glu Val 660 Val Set Gin Val Ile 740 Val Ile Ile Ile ;lu Pal jer 'ys Ala Phe 645 Thr Gin Glu Ala Ala 725 Val Ser Glu Sez Asn P Gin T) Leu E Lys 2 Tyr 630 Trp Leu Arg Asp His 710 Asn Gin Trp Trp Ser sn rp ,ro krg 615 rhr Arg Leu Tyr Va1 695 Thr Phe Ser Ile Lys 775 Sei Leu Gin Phe C 570 Lys Met Tyr 585 Val Pro Asp 600 Leu Asp Gly Val Val Met Ile Ile Asn 650 Trp Lys Pro 665 Val Ile Asn 680 Gly Asn His Val Thr Val Asn Leu Thr 730 Leu Ser Ala 745 Leu Ser Pro 760 Asn Leu Asn *Vai Lys Lys 31n Ile Arg Tyr Gly Leu lu Leu Leu Asp 635 Gly Leu His Thr Leu 715 Phe Tyi Sex Gl Ty: 79.

Val T Cys p Gly 620 Ile Asp Met His Lys 700 Ala Ser Pro Asp i Asp 780 r Tyr 5 lyr ~la 605 Cyr Lys rhr Lys Thr 685 Phe lie Trp Leu Tyr 765 G1l I l Asp 590 Val Trp Va1 Met Asn 670 Ser Thr Asn Pro Asn 750 Lys r Gi.

H i.- 5

A

T

S

P

I

6 ia Lys yr Ala er Asn to Met 640 ,ys Lys ~sp Set :ys Asn ?he Leu Ser Ile 720 Met Set 735 Ser Set Leu Met Ile Lys Asp His 800 Phe Met 815 790 Glu Lys Tyr Gin Phe Set Leu Tyr Pro Ile 805 810 Glu Gly Val Gly Lys Pro Lys Ile 820 Asn Set Phe Thr Gin Asp Asp 830 11.3 Ile Glu Lys His Gin Ser Asp Ala Gly Leu Tyr Val Ile Val Pro Vai 835 840 845 Ile Ile Ser Ser Ser Ile Leu Leu Leu Gly Thr Leu Leu Ile Ser His 5850 855 860 Gin Arg Met Lys Lys Leu Phe Trp Giu Asp Val Pro Asn Pro Lys Asn 865 870 875 880 .0Cys Ser Trp Ala Gin Gly Leu Asn Phe Gin Lys Met Leu Giu Giy Ser 885 890 895 Met Phe Val Lys Ser His His His Ser Leu Ile Ser Ser Thr Gin Gly 900 905 910 His Lys His Cys Gly Arg Pro Gin Gly Pro Leu His Arg Lys Thr Arg 915 920 925 Asp Leu Cys Ser Leu Val Tyr Leu Leu Thr Leu Pro Pro Leu Leu Ser )0930 935 940 Tyr Asp Pro Ala Lys Ser Pro Ser Val Arg Asn Thr Gin Giu Ser Ile 945 950 955 960 5 Lys Lys Lys Lys Lys Lys Leu Giu Giy 965 INFORMATION FOP. SEQ ID.NO:7: 30 SEQUENCE CHARACTERISTICS: LENGTH: 1216 amino acids TYPE: amino acid STRANDEDNESS: single TOPOLOGY: linear (ii) MOLECULE TYPE: protein (xi) SEQUENCE DESCRIPTION: SEQ ID NO:7: Met Ile Cys Gin Lys Phe Cys Val Val Leu Leu His Trp Giu Phe Ile 1 5 10 S Tyr Val Ile Thr Ala Phe Asn Leu Ser Tyr Pro Ile Thr Pro Trp Arg 25 Phe Lys Leu Ser Cys Met Pro Pro Asn Ser Thr Tyr Asp Tyr Phe Leu 3035 40 Leu Pro Ala Gly Leu Ser Lys Asn Th-r Ser Asn Ser Asn Gly His Tyr 55 Glu Asn Arg Ser Ile Glu 145 Leu Gin Cys Leu Pro 225 Leu Trp Tyr Ser Glu 305 Thr Leu Asn Thr Gin 130 Ser Leu Lys Cys Leu 210 Leu Gly Ser Ser Ala 290 Val Ala Ser Cys Val 115 Cys Leu Tyr Gly Glu 195 Met Met Leu Ser Glu 275 Thr Gin Va1 Lys Ser 100 Asn Trp Phe Val Ser 180 Cys Cys Ser His Pro 260 Asn Ser Val Glu Thr Leu Ser Leu Lys Leu 165 Phe Leu Leu Val Met 245 Pro Ser Leu Arg Pro 70 Thr Cys Leu Lys Asn 150 Pro Gin Va1 Lys Gin 230 Glu Leu Thr Leu Gly 310 L1

P]

A

V;

G

1

L

G

M

P

I

2

P

I

V

T

I

114 ys Phe he His la Asp al Phe 120 iy Asp 35 eu Phe lu Val et Val ro Vai 200 le Thr 15 ro Ile ie Thr ral Pro 'hr Val 280 ral Asp !95 .ys Arg Asn Cys Asn 105 Gin Leu Arg Leu His 185 Pro Ser Asn Asp Phe 265 Ile Ser Leu Ser Cys 90 lie Gin Lys Asn Glu 170 Cys Thr Gly Met Asp 250 Pro Arg Ile Asp Ser 75 Phe Glu Ile Leu Tyr 155 Asp Asn Ala Gly Vai 235 Gly Leu Glu Leu Gly 315 Gly Arg Gly Asp Phe Asn Ser Cys Lys Val 220 Lys Asn Gin Ala Pro 300 Pro Thr Ser Lys Ala 125 Ile Tyr Pro Ser Leu 205 Ile Pro Leu Tyr Asp 285 Gly Gly His Glu Thr 110 Asn Cys Lys Leu Val 190 Asn Phe Asp Lys Gin 270 Lys Ser Ile Phe Gin Phe Trp Tyr Val Val 175 His Asp Gin Pro Ile 255 Val Ile Ser Trp Ser Asp Val Asn Val His 160 Pro Glu Thr Ser Pro 240 Ser Lys Val Tyr Ser 320 Asp Trp Ser Thr Pro Arg Val Phe Thr Thr Gin Asp Val 325 330 Ile Tyr Phe 115 Pro Pro Lys Ile Leu Thr Ser Val Gly Ser Asn Val Ser Phe His Cys 340 345 350 Ile Tyr Lys Lys Giu Asn Lys Ile Val Pro Ser Lys Giu Ile Val Trp 355 360 365 Trp Met Asn Leu Ala Giu Lys Ile Pro Gin Ser Gin Tyr Asp Val Val 370 375 380 LO0 Ser Asp His Val Ser Lys Vai Thr Phe Phe Asn Leu Asn Giu Thr Lys 385 390 395 400 Pro Arg Giy Lys Phe Thr Tyr Asp Ala Val Tyr Cys Cys Asn Giu His 405 410 415 Giu Cys His His Arg Tyr Ala Giu Leu Tyr Val Ile Asp Val Asn Ile 420 425 430 Asn Ile Ser Cys Giu Thr Asp Gly Tyr Leu Thr Lys Met Thr Cys Arg 435 440 445 Trp Ser Thr Ser Thr Ile Gin Ser Leu Ala Giu Ser Thr Leu Gin Leu 450 455 460 Arg Tyr His Arg Ser Ser Leu Tyr Cys Ser Asp Ile Pro Ser Ile His 9465 470 475 480 *Pro Ile Ser Giu Pro Lys Asp Cys Tyr 'Leu Gin Ser Asp Gly Phe Tyr 0485 490 495 Giu Cys Ile Phe Gin Pro Ile Phe Leu Leu Ser Giy Tyr Thr Met Trp 500 505 510 Ile Arg Ile Asn His Ser Leu Giy Ser Leu Asp Ser Pro Pro Thr Cys 35 515 520 525 Val Leu Pro Asp Ser Vai Vai Lys Pro Leu Pro Pro Ser Ser Val Lys 99:530 535 540 40 Ala Giu Ile Thr Ile Asn Ile Gly Leu Leu Lys Ile Ser Trp Giu Lys 545 550 55560 Pro Val Phe Pro Giu Asn Asn Leu Gin Phe Gin Ile Arg Tyr Gly Leu 565 570 575 Ser Gly Lys Giu Val Gin Trp Lys Met Tyr Giu Val Tyr Asp Ala Lys 580 -585 590 Ser Lys Ser Val Ser Leu Pro Vai Pro Asp Leu Cys Ala Val Tyr Aia 595 600 605 Val Gin Vai Arg Cys Lys Arg Leu Asp Gly Leu Gly Tyr Trp Ser Asn 610 615 620 T rp 625 Arg Giu Leu Gly Trp 705 Gly Lys Cys Tyr T rp 785 Phe Giu Ile Ile Gin 865 Ser Gly Ly s Cys Thr 690 Thr Al a Val1 Val P he 770 Leu Ile Giy Giu Ile 850 Arg As n Pro As n Se r 675 T rp Glu Ser As n Ile 755 Ile Arg Pro Vali Lys 835 Ser Met Pro Glu Val1 660 Val1 Ser Gin Val1 Ile 740 Val1 Ile Ile Ile Gly 820 His Ser Lys Ala Tyr 630 Phe Trp 645 Thr Leu Gin Arg Giu Asp Ala His 710 Ala Asn 725 Vai Gin Ser Trp Giu Trp Ser Ser 790 Giu Lys 805 Lys Pro Gin Ser Ser Ile Lys Leu 870 116 Thr Val Vz Arg Ile I2 Leu Trp L~ Tyr Vail1 680 Val Giy A~ 695 Thr Val Tt Phe Asn L~ Ser Leu S~ Ile Leu S 760 Lys Asn L 775 Ser Val L' Tyr Gin P1 Lys Ile I.

8; Asp Ala G.

840 Leu Leu L 855 Phe Trp G 1i Met Asp Ile Lys Val Pro Met 635 0* 0 Le is le ~ie 25 ly iu Asn 650 Pro Asn His Val1 Thr 730 Ala Pro As n Lys Ser 810 As n Leu Giy Asp Gly Leu His Thr Leu 715 Phe Tyr Ser Giu Tyr 795 Leu Ser Tyr Thr Vali 875 Asp Met His Ly s 700 Al a Ser Pro Asp Asp 780 Tyr Tyr P he Val Leu 860 Pro Thr Lys Thr 685 P he Ile T rp Leu Tyr 765 Giy Ile Pro Thr Ile 845 Leu Asn Met Lys 655 Asn Asp 670 Ser Cys Thr Phe Asri Ser Pro Met 735 Asn Ser 750 Lys Leu Giu Ile His Asp Ile Phe 815 Gin Asp 830 Vai Pro Ile Ser Pro Lys 640 Lys Ser Asn Le u Ile 720 Ser Ser Met Lys His 800 Met Asp Val1 His As n 880 Cys Ser Trp, Aia Gin Gly Leu Asn Phe Gin Lys Pro Giu Thr 885 890 Phe Giu 895 117 40 0 :6 0 *see 500 His Leu Phe Ile Lys His Thr Ala Ser Val Thr Cys Gly Pro Leu Leu 900 905 910 Leu Glu Pro Giu Thr Ile Ser GJlu Asp Ile Ser Val Asp Thr Ser Trp 915 920 925 Lys Asn Lys Asp Giu Met Met Pro Thr Thr Vai Val Ser Leu Leu Ser 930 935 940 Thr Thr Asp Leu Giu Lys Giy Ser Val Cys Ile Ser Asp Gin Phe Asn 945 950 955 960 Ser Vai Asn Phe Ser Giu Ala Giu Giy Thr Giu Val Thr Tyr Giu Asp 965 970 975 Giu Ser Gin Arg Gin Pro Phe Vai Lys Tyr Ala Thr Leu Ile Ser Asn 980 985 990 Ser Lys Pro Ser Giu Thr Giy Giu Giu Gin Gly Leu Ile Asn Ser Ser 995 1000 1005 Vai Thr Lys Cys Phe Ser Ser Lys Asn Ser Pro Leu Lys Asp Ser Phe 1010 1015 1020 Ser Asn Ser Ser Trp Giu Ile Giu Ala Gin Ala Phe Phe Ile Leu Ser 1025 1030 1035 1040 Asp Gin His Pro Asn Ile Ile Ser Pro His Leu Thr Phe Ser Giu Gly 1045 1050 1055 Leu Asp Giu Leu Leu Lys Leu Giu Gly Asn Phe Pro Giu Giu Asn Asn 1060 1065 1070 Asp Lys Lys Ser Ile Tyr Tyr Leu Gly Val Thr Ser Ile Lys Lys Arg 1075 1080 1085 Giu Ser Gly Val Leu Leu Thr Asp Lys Ser Arg Vai Ser Cys Pro Phe 1090 1095 1100 Pro Ala Pro Cys Leu Phe Thr Asp Ile Arg Vai Leu Gin Asp Ser Cys 1105 1110 1115 .1120 Ser His Phe Val Giu Asn Asn Ile Asn Leu Gly Thr Ser Ser Lys Lys 1125 1130 1135 Thr Phe Ala Ser Tyr Met Pro Gin Phe Gin Thr Cys Ser Thr Gin Thr 1140 1145 1150 His Lys Ile Met Giu Asn Lys Met Cys Asp Leu Thr Val Phe His Arg 1155 1160 1165 Asn Leu Gin Ile Cys Val Ile Met Gly Asn Ile Lys Cys Asn Arg Leu 1170 1175 1180 118 Leu Trp Val Gly Glu Arg Lys Glu Thr Arg Val. Lys Phe Glu Asn Asn 1185 1190 1195 1200 Cys Ser Lys Lys Lys Lys Lys Lys Asn Ser Arg Pro Ala Arg Pro Asp 1205 1210 1215 INFORMATION FOR SEQ ID NO:8: SEQUENCE CHARACTERISTICS: LENGTH: 3599 base pairs TYPE: nucleic acid STRANDEDNESS: single TOPOLOGY: linear (ii) MOLECULE TYPE: cDNA 0 .s (xi) SEQUENCE DESCRIPTION: SEQ ID NO:8: GCGGCCGCCA GTGTGATGGA TATCTGCAGA ATTCGGCTTT

S

GACCCCCGGA

TTTTGTTACA

o CTCCTTGGAG

TGCCTGCTGG

AACCTAAGTT

GTTGCTTTCG

AGACATTTGT

o TACAGTGCTG

AGAATCTATT

TAGAAGATTC

GTGTTCACGA

TCCTTATGTG

o TTCAGCCCAT

TCAAGGTGTA

TTGGGAATTT

ATTTAAGTTG

GCTCTCAAAG

TAATTCAAGT

GAGTGAGCAA

TTCAACAGTA

GCTAAAAGGA

CAGGAAT TAT

ACCTCTGGTT

ATdTTGTGAA

TTTGAAAATC

AAATATGGTG

CTTCTCTGAA

ATTTATGTGA

TCTTGCATGC

AATACTTCAA

GGTACTCACT

GATAGAAACT

AATTCTTTAG

GACTTAAAAT

AACTATAAGG

CCCCAAAAAG

TGTCTTGTGC

ACATCTGGTG

AAGCCTGATC

GTAAGATGAT

TAACTGCGTT

CACCAAATTC

ATTCGAATGG

TTTCTAACTT

GCTCCTTATG

TTTTTCAACA

TATTCATCTG

TCCATCTTTT

GCAGTTTTCA

CTGTGCCAAC

GAGTAATTTT

CACCATTAGG

CTCTGCCTTC

TTGTCAAAAA

TAACTTGTCA

AACCTATGAC

ACATTATGAG

ATCCAAAACA

TGCAGACAAC

AATAGATGCA

TTATGTGGAG

ATATGTTCTG

GATGGTTCAC

AGCCAAACTC

CCAGTCACCT

TTTGCATATG

GGTCGAGTTG

TTCTGTGTGG

TATCCAATTA

TACTTCCTTT

ACAGCTGTTG

ACTTTCCACT

ATTGAAGGAA

AACTGGAACA

TCATTATTTA

CCTGAAGTGT

TGCAATTGCA

AACGACACTC

CTAATGTCAG

GAAATCACAG

120 180 240 300 360 420 480 540 600 660 720 780 840 ATGATGGTA TTTAAAGATT TCTTGGTCCA GCCCACCATT GGTACCATTT CCACTTCAAT 119

ATCAAGTGAA

CAGCTACATC

GGGGCAAGAG

CCACACAAGA

CTTTTCACTG

0

GGATGAATTT

GCAAAGTTAC

CAGTGTACTG

ATGTCAATAT

GGTCALACCAG

:0

GCAGCCTTTA

ATTTGCAGAG

5 ACACA.ATGTG

TCCTTCCTGA

TAAACATTGG

AATTCCAGAT

ATGATGCAAA

35 TTCAGGTGCG

CCTACACAGT

TTAATGGAGA

AAAATGACTC

GAACATGGTC

CACATACTGT

TAACCTTTTC

TAAACAGCAG

ATTTTATTAT

CTTCATCTGT

ATATTCAGAG

CCTGCTAGTA

ACTGGATGGC

TGT CAT AT AC

CATCTATAAG

AGCTGAGAAA

TTTTTTCAAT

CTGCAATGAA

CAATATCTCA

TACAATCCAG

CTGTTCTGAT

TGATGGTTTT

GATTAGGATC

TTCTGTGGTG

ATTATTGAAA

TCGCTATGGT

ATCAAAATCT

CTGTAAGAGG

TGTCATGGAT

TACTATGAAA

ATTGTGCAGT

AGAAGATGTG

TACGGTTCTG

ATGGCCTATG

TTGTGTGATT

TGAGTGGAAA

TAAGAAGTAT

AATTCTACAA

GACAGTATAC

CCAGGAATCT

TTTCCACCTA

AAGGAAAACA

ATTCCTCAAA

CTGAATGA;A

CATGAATGCC

TGTGAAACTG

TCACTTGCGG

ATTCCATCTA

TATGAATGCA

AATCACTCTC

AAGCCACTGC

ATATCTTGGG

TTAAGTGGAA

GTCAGTCTCC

CTAGATGGAC

ATAAAAGTTC.

AAGGAGAAAA

GTTCAGAGAT

GGAAATCACA

GCCATCAATT

AGCAAAGTAA

GTTTCCTGGA

AATCTTAATG

TATATCCATG

CAGTTATCAG

TTCCTGGGTC

GGAGTGACTG

AAATTCTGAC

AGATTGTTCC

GCCAGTATGA

CCAAACCTCG

ATCATCGCTA

ATGGGTACTT

AAAGCACTTT

TTCATCCCAT

TTTTCCAGCC

TAGGTTCACT

CTCCATCCAG

AAAAGCCAGT

AAGAAGTACA

CAGTTCCAGA

TGGGATATTG

CTATGAGAGG

ATGTCACTTT

ATGTGATAAA

CGAAATTCAC

CAATTGGTGC

ATATCGTGCA

TACTATCACC

AAGATGGTGA

ATCATTTTAT

AGAAGC TGAC

TTCGTATGAG

GAGTACTCCT

AAGTGTTGGQ-

CTCAAAAGAG

TGTTGTGAGT

AGGAAAGTTT

TGCTGAATTA

AACTAAAATG

GCAATTGAGG

ATCTGAGCCC

AATCTTCCTA

TGACTCTCCA

TGTGAAAGCA

CTTTCCAGAG

ATGGAAGATG

CTTGTGTGCA

GAGTAATTGG

ACCTGAATTT

ACTTTGGAAG

CCATCATACT

TTTCCTGTGG

TTCTGTTGCA

GTCACTCAGT

CAGTGATTAC

AATAAAATGG

CCCCATTGAG

AAGATTGTCT

GTTCAGGTGA

CGTGTCTTTA

TCTA.ATGTTT

ATTGTTTGGT

GATCATGTTA

ACCTATGATG

TATGTGATTG

ACTTGCAGAT

TATCATAGGA

AAAGATTGCT

TTATCTGGCT

CCAACATGTG

GAAATTACTA

AATAACCTTC

TATGAGGTTT

GTCTATGCTG

AGCAATCCAG

TGGAGAATAA

CCCCTGATGA

TCCTGCAATG

ACAGAGCAAG

AATTTTAATT

GCTTATCCTT

AAGCTAATGT

CTTAGAATCT

AAGTACCAGT

960 1020 1080 1140 1200 1260 1320 1380 1440 1500 1560 1620 1680 1740 1800 1860 1920 1980 2040 2100 2160 2220 2280 2340 2400 2460 2520 120

TCAGTCTTTA

CTCAAGATGA

TTATTTCCTC

AGCTATTTTG

0 TTCAGAAGCC

GTCCTCTTCT

AAAATAAAGA

AAAAGGGTTC

GTACTGAGGT

0 TGATCAGCAA

TCACCAAGTG

GGGAGATAGA

CACACCTCAC

AAGAAAATAA

10 AGAGTGGTGT

TATTCACGGA

ACTTAGGAAC

CCCAATATTT

TAT TGAAAAA

TTCCATCTTA

GGAAGATGTT

AGAAACGTTT

TTTGGAGCCT

TGAGATGATG

TGTTTGTATT

AACCTATGAG

C TCT AAAC CA

CTTCTCTAGC

GGCCCAGGCA

ATTCTCAGAA

TGATAAAAAG

GCTTTTGACT

CATCAGAGTT

TTCTAGTAAG

ATGGAAGGAG

CACCAGAGTG

TTGCTTGGAA

CCGAACCCCA

GAGCATCTTT

GAAACAATTT

CCAACAACTG

AGTGACCAGT

GACGAAAGCC

AGTGAAACTG

AAAAATTCTC

TTTTTTATAT

GGATTGGATG

TCTATCTATT

GACAAGTCAA

CTCCAGGACA

AAGACTTTTG

TGGGAAAACC

ATGCAGGTTT

CATTATTAAT

AGAATTGTTC

TTATCAAGCA

CAGAAGATAT

TGGTCTCTCT

TCAACAGTGT

AGAGACAACC

GTGAAGAACA

CGTTGAAGGA

TATCGGATCA

AACTTTTGAA

ATTTAGGGGT

GGGTATCGTG

GTTGCTCACA

CATCTTACAT

AAAGATAATT

ATATGTAATT

ATCACACCA.A

CTGGGCACAA

TACAGCATCA

CAGTGTTGAT

ACTTTCAACA

TAACTTCTCT

CTTTGTTAAA

AGGGCTTATA

TTCTTTCTCT

GCATCCCAAC

ATTGGAGGGA

CACCTCAATC

CCCATTCCCA

CTTTGTAGAA

GCCTCAATTC

A.ATAGTTTCA

GTGCCAGTAA

AGAATGAAAA

GGACTTAATT

GTGACATGTG

ACATCATGGA

ACAGATCTTG

GAGGCTGAGG

TACGCCACGC

AATAGTTCAG

AATAGCTCAT

ATAATTTCAC

AATTTCCCTG

AAAAAGAGAG

GCCCCCTGTT

AATAATATCA

CAAACTTGTT

2580 2640 2700 2760 2820 2880 2940 3000 3060 3120 3180 3240 3300 3360 3420 3480 3540 3599 CTACTCAGAC TCATAAGATC ATGGAAAACA AGATGTGTGA CCTAACTGTG TAATCTAGA INFORMATION FOR SEQ ID NO:9: SEQUENCE CHARACTERISTICS: LENGTH: 17 base pairs TYPE: nucleic acid STRANDEDNESS: single TOPOLOGY: linear (ii) MOLECULE TYPE: cDNA 121 (xi) SEQUENCE DESCRIPTION: SEQ ID NO:9: NNNNNTACCT TTTCCAG 17 INFORMATION FOR SEQ ID NO:iO: SEQUENCE CHARACTERISTICS: LENGTH: 839 amino acids TYPE: amino acid STRANDEDNESS: single TOPOLOGY: linear (ii) MOLECULE TYPE: protein (xi) SEQUENCE DESCRIPTION: SEQ ID )Met Ile Cys Gln Lys Phe Cys Val Val Leu Leu His Trp Giu Phe Ile 0. 0:1 0.::00Tyr Val Ile Thr Ala Phe Asn Leu Ser Tyr Pro Ile Thr Pro Trp Arg .00..0 0 20 25 Phe Lys Leu Ser Cys Met Pro Pro Asn Ser Thr Tyr Asp Tyr Phe Leu 0 0 35 40 Leu Pro Ala Giy Leu Ser Lys Asn Thr Ser Asn Ser Asn Gly His Tyr )50 55 *0000 Glu Thr Ala Val Giu Pro Lys Phe Asn Ser Ser Giy Thr His Phe Ser 70 75 Asn Leu Ser Lys Thr Thr Phe His Cys Cys Phe Arg Ser Glu Gin Asp 90 :o~oooArg Asn Cys Ser Leu Cys Ala Asp Asn Ilie Glu Gly Lys Thr Phe Vai 0**100 105 110 Ser Thr Val Asn Ser Leu Val Phe Gin Gin Ile Asp Ala Asn Trp Asn 115 120 125 Ile Gin Cys Trp Leu Lys Gly Asp Leu Lys Leu Phe Ile Cys Tyr Val 130 135 140 Giu Ser Leu Phe Lys Asn Leu Phe Arg Asn Tyr Asn Tyr Lys Val His 145 150 155 160 0 Leu Leu Tyr Vai Leu Pro Giu Val Leu Giu Asp Ser Pro Leu Vai Pro 165 170 175 122 Gin Lys Giy Ser Phe Gin Met Val His Cys Asn Cys Ser Val His Giu 180 185 190 Cys Cys Giu Cys Leu Val Pro Val Pro Thr Ala Lys Leu Asn Asp Thr 195 200 205 Leu Leu Met Cys Leu Lys Ile Thr Ser Gly Giy Val Ile Phe Gin Ser 210 215 220 0 Pro Leu Met Ser Val Gin Pro Ile ASn Met Val. Lys Pro Asp Pro Pro 225 230 235 240 Leu Gly Leu His Met Giu Ile Thr Asp Asp Giy Asn Leu Lys Ile Ser 245 250 255 Trp Ser Ser Pro Pro Leu Vai Pro Phe Pro Leu Gin Tyr Gin Vai Lys 260 265 270 Tyr Ser Giu Asn Ser Thr Thr Vai Ile Arg Giu Ala Asp Lys Ile Val ?0275 280 285 0. 0:Ser Ala Thr Ser Leu Leu Vai Asp Ser Ile Leu Pro Giy Ser Ser Tyr 290 295 300 Giu Val Gin Val Arg Giy Lys Arg Leu Asp Gly Pro Gly Ile Trp Ser 305 310 315 320 Asp Trp Ser Thr Pro Arg Vai Phe Thr..Thr Gin Asp Val Ile Tyr Phe 325 330 335 Pro Pro Lys Ile Leu Thr Ser Val Gly Ser Asn Val Ser Phe His Cys 340 345 350 I *le Tyr Lys Lys Giu Asn Lys Ile Val Pro Ser Lys Giu Ile Vai Trp 355 360 365 Trp Met Asn Leu Ala Giu Lys Ile Pro Gin Ser Gin Tyr Asp Val Val 370 375 380 o 40 Ser Asp His Vai Ser Lys Vai Thr Phe Phe Asn Leu Asn Giu Thr Lys 385 390 395 400 Pro Arg Gly Lys Phe Thr Tyr Asp Ala Val Tyr Cys Cys Asn Giu His 405 410 415 Glu Cys His His Arg Tyr Ala Giu Lec Tyr Val Ile Asp Val Asn Ile 420 425 430 Asn Ile Ser Cys Giu Thr Asp Gly Tyr Leu Thr Lys Met Thr Cys Arg 435 440 445 Trp Ser Thr Ser Thr Ile Gin Ser Leu Ala Glu Ser Thr Leu Gin Leu 450 455 460 123 a Arg 465 Pro Glu Ile Val Ala 545 Pro Ser Ser Va1 Trp 625 Arg Glu Leu Gly Trp 705 Gly Tyr Ile Cys Arg Leu 530 Glu Va1 Gly Lys Gin 610 Ser Gly Lys Cys Thr 690 Thr Ala His Ser Ile Ile 515 Pro Ile Phe Lys Ser 595 Val Asn Pro Asn Ser 675 Trp Glu Ser Arg Glu Phe 500 Asn Asp Thr Pro Glu 580 Va1 Arg Pro Glu Val 660 Va1 Ser Gin Va1 Ser Pro 485 Gin His Ser Ile Glu 565 Val Ser Cys Ala Phe 645 Thr Gin Glu Ala Ala 725 Ser 470 Lys Pro Ser Val Asn 550 Asn Gin Leu Lys Tyr 630 Trp Leu Arg Asp His 710 Asn Leu Asp Ile Leu Val 535 Ile Asn Trp Pro Arg 615 Thr Arg Leu Tyr Val 695 Thr Phe Tyr Cys Phe Gly 520 Lys Gly Leu Lys Va1 600 Leu Vai Ile Trp Val 680 Gly Val Asn Cys Tyr Leu 505 Ser Pro Leu Gin Met 585 Pro Asp Val Ile Lys 665 Ile Asn Thr Leu Ser Leu 490 Leu Leu Leu Leu Phe 570 Tyr Asp Gly Met Asn 650 Pro Asn His Va1 Thr 730 Asp 475 Gin Ser Asp Pro Lys 555 Gin Glu Leu Leu Asp 635 Gly Leu His Thr Leu 715 Phe Ile Ser Gly Ser Pro 540 Ile Ile Val Cys Gly 620 Ile Asp Met His Lys 700 Ala Ser Pro Asp Tyr Pro 525 Ser Ser Arg Tyr Ala 605 Tyr Lys Thr Lys Thr 685 Phe Ile Trp Ser Gly Thr 510 Pro Ser Trp Tyr Asp 590 Val Trp Val Met Asn 670 Ser Thr Asn Pro Ile Phe 495 Met Thr Va1 Glu Gly 575 Ala Tyr Ser Pro Lys 655 Asp Cys Phe Ser Met 735 His 480 Tyr Trp Cys Lys Lys 560 Leu Lys Ala Asn Met 640 Lys Ser Asn Leu Ile 720 Ser 124 Lys Val Asn Cys Val Ile 755 Tyr Phe Ile 770 Ile 740 Val1 Val Gin Ser Leu Ala Tyr Pro Leu Ser Trp Ile Pro Ser Asp Tyr 765 Giy Asn Ser Ser 750 Lys Leu Met Glu Ile Lys Ile Glu Trp Lys 775 Leu Asn Giu Asp 780 Tyr Trp 785 P he Leu Arg Ile Ser Ser 790 Ser Val Lys Lys Tyr 795 Leu Ile His Asp His 800 Ile Pro Ile Giu 805 Lys Lys Tyr Gin Phe Ser 810 As n Tyr Pro Ile Phe Met 815 Asp Asp Giu Gly Vai Ile Giu Lys 835 Pro Lys Ile Ser Phe Thr Gin 830 Gin Ser Asp 0:.

INFORMATION FOR SEQ ID NO:ii: SEQUENCE CHARACTERISTICS: LENGTH: 2624 base pairs TYPE: nucleic acid STRANDEDNESS: single TOPOLOGY: linear (ii) MOLECULE TYPE: cDNA 35 (xi) SEQUENCE DESCRIPTION: SEQ ID NO:i1: GCGGCCGCCA GTGTGATGGA TATCTGCAGA ATTCGGCTTT GACCCCCGGA TCAAGGTGTA CTTCTCTGAA GTAAGATGAT TTTTGTTACA TTGGGAATTT ATTTATGTGA TAACTGCGTT CTCCTTGGAG ATTTAAGTTG TCTTGCATGC CACCAAATTC TGCCTGCTGG GCTCTCAAAG AATACTTCAA ATTCGAATGG AACCTAAGTT TAATTCAAGT GGTACTCACT TTTCTAACTT GTTGCTTTCG GAGTGAGCAA GATAGAAACT GCTCCTTATG AGACATTTGT TTCAACAGTA AATTCTTTAG TTTTTCAACA TACAGTGCTG GCTAAAAGGA GACTTAAAAT TATTCATCTG

CTCTGCCTTC

TTGTCAAAAA

TAACTTGTCA

AACCTATGAC

ACATTATGAG

ATCCAAAACA

TGCAGACAAC

AATAGATGCA

TTATGTGGAG

GGTCGAGTTG

TTCTGTGTGG

TATCCAATTA

TACTTCCTTT

ACAGCTGTTG

ACTTTCCACT

ATTGAAGGAA

AACTGGAACA

TCATTATTTA

120 180 240 300 360 420 480 540 125

V.

*.VVV.

V

V.

V.

V

AGAATCTATT

TAGAAGATTC

GTGTTCACGA

TCCTTATGTG

TTCAGCCCAT

ATGATGGTAA

ATCAAGTGAA

CAGCTACATC

GGG4GCAAGAG J CCACACA.AGA

CTTTTCACTG

GGATGAATTT

GCAAAGTTAC

CAGTGTACTG

o ATGTCAATAT

GGTCAACCAG

GCAGCCTTTA

ATTTGCAGAG

ACACAATGTG

o TCCTTCCTGA

TAAACATTGG

AATTCCAGAT

ATGATGCAAA

TTCAGGTGCG

o CCTACACAGT

CAGGAATTAT

ACCTCTGGTT

ATGTTGTGAA

TTTGAAAATC

AAATATGGTG

TTTAAAGATT

ATATTCAGAG

CCTGCTAGTA

ACTGGATGGC

TGTCATATAC

CATCTATAAG

AGCTGAGAAA

TTTTTTCAAT

CTGCAATGAA

CAATATCTCA

TACAATCCAG

CTGTTCTGAT

TGATGGTTTT

GATTAGGATC

TTCTGTGGTG

ATTATTGAAA

TCGCTATGGT

ATCAAAATCT

CTGTAAGAGG

TGTCATGGAT

AACTATALAGG

C CC CAAAAAG

TGTCTTGTGC

ACATCTGGTG

AAGCCTGATC

TCTTGGTCCA

AATTCTACA.A

GACAGTATAC

CCAGGAATCT

TTTCCACCTA

AAGGAAAACA

ATTCCTCAAA

CTGAATGAAA

CATGAATGCC

TGTGAAACTG

TCACTTGCGG

ATTCCATCTA

TATGAATGCA

AATCACTCTC

AAGCCACTGC

ATATCTTGGG

TTAAGTGGAA

GTCAGTCTCC

CTAGATGGAC

ATAAAAGTTC

TCCATCTTTT

GCAGTTTTCA

CTGTGCCAAC

GAGTAATTTT

CACCATTAGG

GCCCACCATT

CAGTTATCAG

TTCCTGGGTC

GGAGTGACTG

AAATTCTGAC

AGATTGTTCC

GCCAGTATGA

CCAAACCTCG

ATCATCGCTA

ATGGGTACTT

AAAGCACTTT

TTCATCCCAT

TTTTCCAGCC

TAGGTTCACT

CTCCATCCAG

AAA.AGCCAGT

AAGAAGTACA

CAGTTCCAGA

TGGGATATTG

CTATGAGAGG

ATATGTTCTG

GATGGTTCAC

AGCCAAACTC

CCAGTCACCT

TTTGCATATG

GGTACCATTT

AGA.AGCTGAC

TTCGTATGAG

GAGTACTCCT

AAGTGTTGGG

CTCAAA.AGAG

TGTTGTGAGT

AGGAAAGTTT

TGCTGAATTA

AACTAAAATG

GCAATTGAGG

ATCTGAGCCC

AATCTTCCTA

TGACTCTCCA

TGTGAAAGCA

CTTTCCAGAG

ATGGAAGATG

CTTGTGTGCA

GAGTAATTGG

ACCTGAATTT

CCTGAAGTGT

TGCAATTGCA

AACGACACTC

CTAATGTCAG

GAAATCACAG

CCACTTCAAT

AAGA'TTGTCT

GTTCAGGTGA

CGTGTCTTTA

TCTAATGTTT

ATTGTTTGGT

GATCATGTTA

ACCTATGATG

TATGTGATTG

ACTTGCAGAT

TATCATAGGA

AAAGATTGCT

TTATCTGGCT

CCAACATGTG

GAAATTACTA

AATAACCTTC

TATGAGGTTT

GTCTATGCTG

AGCAATCCAG

TGGAGAATAA

600 660 720 780 840 900 960 1020 1080 1140 1200 1260 1320 1380 1440 1500 1560 1620 1680 1740 1800 1860 1920 1980 2040 2100 TTAATGGAGA TACTATGAAA AAGGAGAAAA ATGTCACTTT ACT TTGGAAG CCCCTGATGA 126 AAAATGACTC ATTGTGCAGT GTTCAGAGAT ATGTGATAAA GAACATGGTC AGAAGATGTG GGAAATCACA CGAAATTCAC CACATACTGT TACGGTTCTG GCCATCAATT CAATTGGTGC TAACCTTTTC ATGGCCTATG AGCAAAGTAA ATATCGTGCA TAAACAGCAG TTGTGTGATT GTTTCCTGGA TACTATCACC 0 ATTTTATTAT TGAGTGGAAA AATCTTAATG AAGATGGTGA CTTCATCTGT TAAGAAGTAT TATATCCATG ATCATTTTAT TCAGTCTTTA CCCAATATTT ATGGAAGGAG TGGGAAAACC CTCA.AGATGA TATTGAAAAA CACCAGAGTG ATTGATAAGG INFORMATION FOR SEQ ID NO:12: SEQUENCE CHARACTERISTICS: LENGTH: 2948 base pairs TYPE: nucleic acid STRANDEDNESS: single TOPOLOGY: linear (ii) MOLECULE TYPE: cDNA

CCATCATACT

TTTCCTGTGG

TTCTGTTGCA

GTCACTCAGT

CAGTGATTAC

AATA.AAATGG

CCCCATTGAG

AAAGATAATT

ATCC

TCCTGCAATG

ACAGAGCAAG

AATTTTAATT

GCTTATCCTT

AAGCTAATGT

CTTAGAATCT

AAGTACCAGT

AATAGTTTCA

2160 2220 2280 2340 2400 2460 2520 2580 2624 (xi) SEQUENCE.DESCRIPTION: SEQ ID NO:12: CCATTGAAGT CAATGGGAGT TTGTTTTGGC ACCA.AAATCA ACGGGGATTT CCAAAATGTC

GTAATAACCC

TAAGCAGAGC

10 AGCTTGCTAG

GTCGAGTTGG

TCTGTGTGGT

ATCCAATTAC

ACTTCCTTTT

0 CAGCTGTTGA

CTTTCCACTG

CGCCCCGTTG

TCGTTTAGTG

CGGCCGCCAG

ACCCCCGGAT

TTTGTTACAT

TCCTTGGAGA

GCCTGCTGGG

ACCTAAGTTT

TTGCTTTCGG

ACGCAAATGG

AACCGTCAGA

TGTGATGGAT

CAAGGTGTAC

TGGGAATTTA

TTTAAGTTGT

CTCTCAAAGA

AATTCAAGTG

AGTGAGCA.AG

GCGGTAGGCG

TCTCTAGAAG

ATCTGCAGAA

TTCTCTGAAG

TTTATGTGAT

CTTGCATGCC

ATACTTCAAA

GTACTCACTT

ATAGAAACTG

TGTACGGTGG

CTGGGTACCA

TTCGGCTTTC

TAAGATGATT

AACTGCGTTT

ACCAAATTCA

TTCGAATGGA

TTCTAACTTA

CTCCTTATGT

GAGGTCTATA

GCTGCTAGCA

TCTGCCTTCG

TGTCAAAAAT

AACTTGTCAT

ACCTATGACT

CATTATGAGA

TCCAAAACAA

GCAGACAACA

120 180 240 300 360 420 480 540 600 127 0 0 .00.

TTGAAGGAAA

ACTGGAACAT

CATTATTTAA

CTGAAGTGTT

GCAATTGCAG

ACGACACTCT

TAATGTCAGT

AAATCACAGA

CACTTCAATA

AGATTGTCTC

TTCAGGTGAG

GTGTCTTTAC

CTAATGTTTC

TTGTTTGGTG

ATCATGTTAG

CCTATGATGC

ATGTGATTGA

CTTGCAGATG

ATCATAGGAG

AAGATTGCTA

TATCTGGCTA

CAACATGTGT

3 AAATTACTAT

ATAACCTTCA

ATGAGGTTTA

TCTATGCTGT

GCAATCCAGC

GACATTTGTT

ACAGTGCTGG

GAATCTATTC

AGAAGATTCA

TGTTCACGA.A

CCTTATGTGT

TCAGCCCATA

TGATGGTAAT

TCA.AGTGAAA

AGCTACATCC

GGGCAAGAGA

CACACAAGAT

TTTTCACTGC

GATGAATTTA

CAAAGTTACT

AGTGTACTGC

TGTCA.ATATC

GTCAACCAGT

CAGCCTTTAC

TTTGCAGAGT

CACAATGTGG

CCTTCCTGAT

AAACATTGGA

ATTCCAGATT

TGATGCAAAA

TCAGGTGCGC

CTACACAGTT

TCAACAGTAA

CTAAAAGGAG

AGGAATTATA

CCTCTGGTTC

TGTTGTGAAT

TTGAAAATCA

AATATGGTGA

TTAAAGATTT

TATTCAGAGA

CTGCTAGTAG

CTGGATGGCC

GTCATATACT

ATCTATAAGA

GCTGAGAAAA

TTTTTCAATC

TGCAATGAAC

AATATCTCAT

ACAATCCAGT

TGTTCTGATA

GATGGTTTTT

ATTAGGATCA

TCTGTGGTGA

TTATTGAAAA

CGCTATGGTT

TCAAAATCTG

TGTAAGAGGC

GTCATGGATA

ATTCTTTAGT

ACTTAAAATT

ACTATAAGGT

CCCAAAAAGG

GTCTTGTGCC

CATCTGGTGG

AGCCTGATCC

CTTGGTCCAG

ATTCTACAAC

ACAGTATACT

CAGGAATCTG

TTCCACCTAA

AGGAAAACAA

TTCCTCAAAG

TGAATGAAAC

ATGAATGCCA

GTGAAACTGA

CACTTGCGGA

TTCCATCTAT

ATGAATGCAT

ATCACTCTCT

AGCCACTGCC

TATCTTGGGA

TAAGTGGAAA

TCAGTCTCCC

TAGATGGACT

TAAAAGTTCC

TT-TTCAACAA

ATTCATCTGT

CCATCTTTTA

CAGTTTTCAG

TGTGCCAACA

AGTAATTTTC

ACCATTAGGT

CCCACCATTG

AGTTATCAGA

TCCTGGGTCT

GAGTGACTGG

AATTCTGACA

GATTGTTCCC

CCAGTATGAT

CAAACCTCGA

TCATCGCTAT

TGGGTACTTA

AAGCACTTTG

TCATCCCATA

TTTCCAGCCA

AGGTTCACTT

TCCATCCAGT

AAAGCCAGTC

AGAAGTACAA

AGTTCCAGAC

GGGATATTGG

TATGAGAGGA

ATAGATGCAA

TATGTGGAGT

TATGTTCTGC

ATGGTTCACT

GCCAAACTCA

CAGTCACCTC

TTGCATATGG

GTACCATTTC

GAAGCTGACA

TCGTATGAGG

AGTACTCCTC

AGTGTTGGGT

TCAAAAGAGA

GTTGTGAGTG

GGAAAGTTTA

GCTGAATTAT

ACTAAAATGA

CAATTGAGGT

TCTGAGCCCA

ATCTTCCTAT

GACTCTCCAC

GTGAAAGCAG

TTTCCAGAGA

TGGAAGATGT

TTGTGTGCAG

AGTAATTGGA

CCTGAATTTT

660 720 780 840 900 960 1020 1080 1140 1200 1260 1320 1380 1440 1500 1560 1620 1680 1740 1800 1860 1920 1980 2040 2100 2160 2220 128

GGAGAATAAT

CCCTGATGAA

CCTGCA.ATGG

CAGAGCAAGC

ATTTTAATTT

CTTATCCTTT

AGCTAATGTA

TTAGAATCTC

AGTACCAGTT

JATAGTTTCAC

ACGATGACAA

ACTAGAATGC

GTAACCAT

TAATGGAGAT

A-AATGACTCA

AACATGGTCA

ACATACTGTT

AACCTTTTCA

AAACAGCAGT

TTTTATTATT

TTCATCTGTT

CAGTCTTTAC

TCAAGATGAT

GTAGGGATCC

AGTGAAAAAA

ACTATGAAA.A

TTGTGCAGTG

GAAGATGTGG

ACGGTTCTGG

TGGCCTATGA

TGTGTGATTG

GAGTGGAAAA

AAGAAGTATT

CCA.ATATTTA

ATTGAAAAAC

AGACATGATA

ATGCTTTATT

AGGAGAAAAA

TTCAGAGATA

GAAATCACAC

CCATCAATTC

GCAAAGTAAA

TTTCCTGGAT

ATCTTAATGA

ATATCCATGA

TGGAAGGAGT

ACCAGAGTGA

AGATACATTG

TGTGAAATTT

TGTCACTTTA

TGTGATAAAC

GAAATTCACT

AATTGGTGCT

TATCGTGCAG

ACTATCACCC

AGATGGTGA.A

TCATTTTATC

GGGAAAACCA

TGCAGGTGAC

ATGAGTTTGG

GTGATGCTAT

CTTTGGAAGC

CATCAT ACT T

TTCCTGTGGA

TCTGTTGCAA

TCACTCAGTG

AGTGATTACA

ATAAAATGGC

CCCATTGAGA

AAGATAATTA

TACAAGGACG

ACAACCCACA

TGCTTTATTT

2280 2340 2400 2460 2520 2580 2640 2700 2760 2820 2880 2940 2948

C

C

C

C.

C.

INFORMATION FOR SEQ ID NO:13: SEQUENCE CHARACTERISTICS: LENGTH: 804 amino acids TYPE: amino acid STRANDEDNESS: single TOPOLOGY: linear (ii) MOLECULE TYPE: protein (xi) SEQUENCE DESCRIPTION: SEQ ID NO:l3: Met Ile Cys Gln Lys Phe Cys Val Val Leu Leu 1 5 10 Tyr Val Ile Thr Ala Phe Asn Leu Ser Tyr Pro 25 Phe Lys Leu Ser Cys Met Pro Pro Asn Ser Thr 40 Leu Pro Ala Gly Leu Ser Lys Asn Thr Ser Asn 55 His Trp Glu Phe Ile Ile Thr Pro Trp Arg Tyr Asp Tyr Phe Leu Ser Asn Gly His Tyr 129 Giu Thr Ala Val Giu Pro Lys Phe Asn Ser Ser Gly Thr His Phe Ser 70 75 Asn Leu Ser Lys Thr Thr Phe His Cys Cys Phe Arg Ser Giu Gin Asp 90 Arg Asri Cys Ser Leu Cys Ala Asp Asn Ile Giu Gly Lys Thr Phe Val 100 105 110 0 Ser Thr Val Asn Ser Leu Val Phe Gin Gin Ile Asp Ala Asn Trp Asn 115 120 125 Ile Gin Cys Trp Leu Lys Giy Asp Leu Lys Leu Phe Ile Cys Tyr Val 130 135 140 Giu Ser Leu Phe Lys Asn Leu Phe Arg Asn Tyr Asn Tyr Lys Val His 145 150 155 160 o Leu Leu Tyr Val Leu Pro Giu Val Leu Giu Asp Ser Pro Leu Val Pro 165 170 175 Gin Lys Giy Ser Phe Gin Met Val His Cys Asn Cys Ser Val His Giu :*.180 185 190 *Cys Cys Giu Cys Leu Val Pro Val Pro Thr Ala Lys Leu Asn Asp Thr 00i 95 200 205 Leu Leu Met Cys Leu Lys Ile Thr Ser Gly Gly Vai Ile Phe Gin Ser 210 215 220 Pro Leu Met Ser Val Gin Pro Ile Asn Met Val Lys Pro Asp Pro Pro 225 230 235 240 Leu Gly Leu His Met Giu Ile Thr Asp Asp Gly Asn Leu Lys Ile Ser 245 250 255 Trp Ser Ser Pro Pro Leu Val Pro Phe Pro Leu Gin Tyr Gin Val Lys 260 265 270 Tyr Ser Giu Asn Ser Thr Thr Val Ile Arg Giu Ala Asp Lys Ile Vai 275 280 285 Ser Ala Thr Ser Leu Leu Vai Asp Ser Ile Leu Pro Gly Ser Ser Tyr 290 295 300 Giu Vai Gin Val Arg Giy Lys Arg Leu Asp Giy Pro Giy Ile Trp Ser 305 310 315 320 0 Asp Trp Ser Thr Pro Arg Vai Phe Thr Thr Gin Asp Val Ile Tyr Phe 325 330 335 130 Pro Pro Lys Ile Leu Thr Ser Val. Gly Ser Asn Val Ser Phe His Cys 340 345 350 Ile Tyr Lys Lys Giu Asn Lys Ile Val Pro Ser Lys Giu Ile Val Trp 355 360 365 Trp Met Asn Leu Ala Giu Lys Ile Pro Gin Ser Gin Tyr Asp Val Val 370 375 380 Ser Asp His Val Ser Lys Val Thr Phe Phe Asn Leu Asn Giu Thr Lys 385 390 395 400 Pro Arg Gly Lys Phe Thr Tyr Asp Ala Val Tyr Cys Cys Asn Giu His 405 410 415 Giu Cys His His Arg Tyr Ala Giu Leu Tyr Vai Ile Asp Val Asn Ile 420 425 430 Asn Ile Ser Cys Giu Thr Asp Giy Tyr Leu Thr Lys Met Thr Cys Arg )435 440 445 Trp Ser Thr Ser Thr Ile Gin Ser Leu Ala Giu Ser Thr Leu Gin Leu 450 455 460 *Arg Tyr His Arg Ser Ser Leu Tyr Cys Ser Asp Ile Pro Ser Ile His **465 470 475 480 **Pro Ile Ser Giu Pro Lys Asp Cys Tyr Leu Gin Ser Asp Giy Phe Tyr 485 490 495 *Giu Cys Ile Phe Gin Pro Ile Phe Leu Leu Ser Gly Tyr Thr Met Trp 500 505 510 Ile Arg Ile Asn His Ser Leu Giy Ser Leu Asp Ser Pro Pro Thr Cys 5515 520 525 Val Leu Pro Asp Ser Vai Val Lys Pro Leu Pro Pro Ser Ser Vai Lys 530 535 540 0 Ala Giu Ile Thr Ile Asn Ile Gly Leu Leu Lys Ile Ser Trp, Giu Lys 545 550 55560 Pro Val Phe Pro Glu Asn Asn Leu Gin Phe Gin Ile Arg Tyr Gly Leu 565 570 575 Ser Gly Lys Giu Val Gin Trp, Lys Met Tyr Giu Val Tyr Asp Ala Lys 580 585 590 Ser Lys Ser Val Ser Leu Pro Val Pro Asp Leu Cys Ala Vai Tyr Ala 0 595 600 605 Val Gin Vai Arg Cys Lys Arg Leu Asp Giy Leu Gly Tyr Trp Ser Asn 610 615 620 131 Trp Ser Asn Pro Ala Tyr Thr Val Val Met Asp Ile Lys Val pro Met 625 630 635 640 Arg Gly Pro Giu Phe Trp Arg Ile Ile Asn Gly Asp Thr Met Lys Lys 645 650 655 Glu Lys Asn Val Thr Leu Leu Trp Lys Pro Leu Met Lys Asn Asp Ser 660 665 670 Leu Cys Ser Val Gin Arg Tyr Val Ile Asn His His Thr Ser Cys Asn 675 680 685 Gly Thr Trp Ser Glu Asp Vai Giy Asn His Thr Lys Phe Thr Phe Leu 690 695 700 Trp Thr Giu Gin Ala His Thr Vai Thr Val Leu Ala Ile Asn Ser Ile 705 710 715 720 Gly Ala Ser Val Ala Asn Phe Asn Leu Thr Phe Ser Trp Pro Met Ser 725 730 735 Lys Val Asn Ile Val Gin Ser Leu Ser Ala Tyr Pro Leu Asn Ser Ser .740 745 750 Cys Vai Ile Val Ser Trp Ile Leu Ser Pro Ser Asp Tyr Lys Leu Met 755 760 765 Tyr Phe Ile Ile Giu Trp Lys Asn Leu A sn Giu Asp Gly Glu Ile Lys J770 775 780 *Trp Leu Arg Ile Ser Ser Ser Val Lys Lys Tyr Tyr Ile His Gly Lys 785 790 795 800 ThrleLeu INFORMATION FOR SEQ ID NO:i4: 0 Ci) SEQUENCE CHARACTERISTICS: CA) LENGTH: 2507 base pairs TYPE: nucleic acid STRANDEDNESS: single TOPOLOGY: linear (ii) MOLECULE TYPE: cDNA 132 (xi) SEQUENCE DESCRIPTION: SEQ ID NO:14: GCGGCCGCCA GTGTGATGGA TATCTGCAGA ATTCGGCTTT CTCTGCCTTC GGTCGAGTTG

S.

S

S

S

S

S.

S

S

GACCCCCGGA TCAAGGTGTA TTTTGTTACA TTGGGAATTT CTCCTTGGAG ATTTAAGTTG 0 TGCCTGCTGG GCTCTCAAAG AACCTAAGTT TAATTCAAGT GTTGCTTTCG GAGTGAGCAA AGACATTTGT TTCAACAGTA TACAGTGCTG GCTAAAAGGA 0 AGAATCTATT CAGGAATTAT TAGAAGATTC ACCTCTGGTT 5 GTGTTCACGA ATGTTGTGAA TCCTTATGTG TTTGAAAATC TTCAGCCCAT AAATATGGTG 0 ATGATGGTAA TTTAAAGATT ATCAAGTGAA ATATTCAGAG 5 CAGCTACATC CCTGCTAGTA GGGGCAAGAG ACTGGATGGC CCACACAAGA TGTCATATAC 0 CTTTTCACTG CATCTATAAG GGATGAATTT AGCTGAGAAA GCAAAGTTAC TTTTTTCAAT CAGTGTACTG CTGCAATGAA ATGTCA.ATAT CAATATCTCA GGTCAACCAG TACAATCCAG

CTTCTCTGAA

ATTTATGTGA

TCTTGCATGC

AATACTTCAA

GGTACTCACT

GATAGAAACT

AATTCTTTAG

GACTTAAAAT

A.ACTATAAGG

CCC CAAAAAG

TGTCTTGTGC

AC AT C TGGTG

AAGCCTGATC

TCTTGGTCCA

AATTCTACAA

GACAGTATAC

CCAGGAATCT

TTTCCACCTA

AAGGAAAACA

ATTCCTCAAA

CTGAATGAAA

CATGAATGCC

TGTGAAACTG

TCACTTGCGG

GTAAGATGAT

TAACTGCGTT

CACCAAATTC

ATTCGAATGG

TTTCTAACTT

GCTCCTTATG

TTTTTCAACA

TATTCATCTG

TCCATCTTTT

GCAGTTTTCA

CTGTGCCAAC

GAGTAATTTT

CACCATTAGG

GCCCACCATT

CAGTTATCAG

TTCCTGGGTC

GGAGTGACTG

AAATTCTGAC

AGATTGTTCC

GCCAGTATGA

CCAAACCTCG

ATCATCGCTA

ATGGGTACTT

AAAGCACTTT

TTGTCAAAAA

TA.ACTTGTCA

AACCTATGAC

ACATTATGAG

ATCCAAAACA

TGCAGACAAC

AATAGATGCA

TTATGTGGAG

ATATGTTCTG

GATGGTTCAC

AGCCAAACTC

CCAGTCACCT

TTTGCATATG

GGTACCATTT

AGAAGCTGAC

TTCGTATGAG

GAGTACTCCT

AAGTGTTGGG

CTCAAAAGAG

TGTTGTGAGT

AGGAAAGTTT

TGCTGAATTA

AACTAAAATG

GCAATTGAGG

TTCTGTGTGG

TATCCAATTA

TACTTCCTTT

ACAGCTGTTG

ACTTTCCACT

ATTGAAGGAA

AACTGGAACA

TCATTATTTA

CCTGAAGTGT

TGCAATTGCA

AACGACACTC

CTAATGTCAG

GAAATCACAG

CCACTTCAAT

AAGATTGTCT

GTTCAGGTGA

CGTGTCTTTA

TCTAATGTTT

ATTGTTTGGT

GATCATGTTA

ACCTATGATG

TATGTGATTG

ACTTGCAGAT

TATCATAGGA

120 180 240 300 360 420 480 540 600 660 720 780 840 900 960 1020 1080 1140 1200 1260 1320 1.380 1440 1500 1560 .GCAGCCTTTA CTGTTCTGAT ATTCCATCTA TTCATCCCAT ATCTGAGCCC AAAGATTGCT 133

ATTTGCAGAG

ACACAATGTG

TCCTTCCTGA

TAAACATTGG

J AATTCCAGAT

ATGATGCAAA

TTCAGGTGCG

CCTACACAGT

TTAATGGAGA

3 AAAATGACTC

GAACATGGTC

CACATACTGT

TAACCTTTTC

TAAACAGCAG

o ATTTTATTAT

TGATGGTTTT

GATTAGGATC

TTCTGTGGTG

ATTATTGAAA

TCGCTATGGT

ATCAAAATCT

CTGTAAGAGG

TGTCATGGAT

TACTATGAAA

ATTGTGCAGT

AGAAGATGTG

TACGGTTCTG

ATGGCCTATG

TTGTGTGATT

TGAGTGGAAA

TATGAATGCA

AATCACTCTC

AAGCCACTGC

ATATCTTGGG

TTAAGTGGAA

GTCAGTCTCC

CTAGATGGAC

ATAAAAGTTC

A.AGGAGAAAA

GTTCAGAGAT

GGAAATCACA

GCCATCAATT

AGCAAAGTAA

GTTTCCTGGA

AATCTTAATG

TTTTCCAGCC AATCTTCCTA TTATCTGGCT

TAGGTTCACT

CTCCATCCAG

AAAAGCCAGT

AAGAAGTACA

CAGTTCCAGA

TGGGATATTG

CTATGAGAGG

ATGTCACTTT

ATGTGATAAA

CGAAATTCAC

CAATTGGTGC

ATATCGTGCA

TACTATCACCt

AAGATGGTGA

GTA.AGTTTAC

TGACTCTCCA

TGTGAAAGCA

CTTTCCAGAG

ATGGAAGATG

CTTGTGTGCA

GAGTAATTGG

ACCTGAATTT

ACTTTGGAAG

CCATCATACT

TTTCCTGTGG

TTCTGTTGCA

GTCACTCAGT

CAGTGATTAC

AATAAAATGG

TATACTT

CCA.ACATGTG

GAAATTACTA

XATAACCTTC

TATGAGGTTT

GTCTATGCTG

AGCAATCCAG

TGGAGAATAA

CCCCTGATGA

TCCTGCAATG

ACAGAGCAAG

AATTTTAATT

GCTTATCCTT

AAGCTAATGT

CTTAGAATCT

1620 1680 1740 1800 1860 1920 1980 2040 2100 2160 2220 2280 2340 2400 2460 2507 a 0* S.

S

*00*o CTTCATCTGT TAAGAAGTAT TATATCCATG INFORMATION FOR SEQ ID Ci) SEQUENCE CHARACTERISTICS: LENGTH: 29 base pairs TYPE: nucleic acid STRANDEDNESS: single TOPOLOGY: linear (ii) MOLECULE TYPE: cDNA (xi) SEQUENCE DESCRIPTION: SEQ ID GTAAGTTATT TGNNNNNATA TCCTAACAG 134 INFORMATION FOR SEQ ID NO:16: SEQUENCE CHARACTERISTICS: LENGTH: 29 base pairs TYPE: nucleic acid STRANDEDNESS: single TOPOLOGY: linear (ii) MOLECULE TYPE: cDNA 3 (xi) SEQUENCE DESCRIPTION: SEQ ID NO:16: GTAAGCATTA GCNNNNNTTT TAAATTCAG 29 INFORMATION FOR SEQ ID NO:17: 0 SEQUENCE CHARACTERISTICS: LENGTH: 29 base pairs TYPE: nucleic acid STRANDEDNESS: single ooo, TOPOLOGY: linear

S

(ii) MOLECULE TYPE: cDNA 0 0 (xi) SEQUENCE DESCRIPTION: SEQ ID NO:17: 0 GTAAGTACCA AANNNNNTTT TCAATATAG 29 5 INFORMATION FOR SEQ ID NO:18: oooo SEQUENCE CHARACTERISTICS: LENGTH: 29 base pairs TYPE: nucleic acid 0 STRANDEDNESS: single o TOPOLOGY: linear (ii) MOLECULE TYPE: cDNA (xi) SEQUENCE DESCRIPTION: SEQ ID NO:18: 0 GTAAGTTATG CANNNNNTTT TTCCTTAAG 29 135 INFORMATION FOR SEQ ID NO:19: SEQUENCE CHARACTERISTICS: LENGTH: 28 base pairs TYPE: nucleic acid STRANDEDNESS: single TOPOLOGY: linear (ii) MOLECULE TYPE: cDNA 0 (xi) SEQUENCE DESCRIPTION: SEQ ID NO:19: GTAAGTATAT TTNNNNAATA TTTAACAG 28 INFORMATION FOR SEQ ID 0 SEQUENCE CHARACTERISTICS: LENGTH: 29 base pairs TYPE: nucleic acid STRANDEDNESS: single TOPOLOGY: linear :5 (ii) MOLECULE TYPE: cDNA ;o (xi) SEQUENCE DESCRIPTION: SEQ ID GTAGGTTATG TANNNNNCCC TCATTACAG 29 INFORMATION FOR SEQ ID NO:21: SEQUENCE CHARACTERISTICS: LENGTH: 29 base pairs TYPE: nucleic acid STRANDEDNESS: single TOPOLOGY: linear (ii) MOLECULE TYPE: DNA (xi) SEQUENCE DESCRIPTION: SEQ ID NO:21: GTAAGAAAAC AGNNNNNTGT TTCAAATAG 2 GTAAGAAAAC AGNNNNNTGT TTCAAATAG 136 INFORMATION FOR SEQ ID NO:22: SEQUENCE CHARACTERISTICS: LENGTH: 29 base pairs TYPE: nucleic acid STRANDEDNESS: single TOPOLOGY: linear (ii) MOLECULE TYPE: cDNA (xi) SEQUENCE DESCRIPTION: SEQ ID NO:22: GTACGTATTA TTNNNNNTAT CTTTTAAAG 29 INFORMATION FOR SEQ ID NO:23: 3 SEQUENCE CHARACTERISTICS: LENGTH: 29 base pairs TYPE: nucleic acid STRANDEDNESS: single TOPOLOGY: linear (ii) MOLECULE TYPE: cDNA 0 (xi) SEQUENCE DESCRIPTION: SEQ ID NO:23: GTATGTCAAG CTNNNNNAAA AATTTCTAG 29 5 INFORMATION FOR SEQ ID NO:24: SEQUENCE CHARACTERISTICS: LENGTH: 29 base pairs TYPE: nucleic acid 0 STRANDEDNESS: single TOPOLOGY: linear (ii) MOLECULE TYPE: cDNA (xi) SEQUENCE DESCRIPTION: SEQ ID NO:24: 0 GTACCTTTTA CTNNNNNCTT ATTTTACAG 29 137 INFORMATION FOR SEQ ID SEQUENCE CHARACTERISTICS: LENGTH: 29 base pairs TYPE: nucleic acid STRANDEDNESS: single TOPOLOGY: linear (ii) MOLECULE TYPE: cDNA 0 (xi) SEQUENCE DESCRIPTION: SEQ ID GTCTGCAGAG ATNNNNNGTC ATTTTGCAG 29 INFORMATION FOR SEQ ID NO:26: 0 SEQUENCE CHARACTERISTICS: LENGTH: 29 base pairs TYPE: nucleic acid STRANDEDNESS: single TOPOLOGY: linear (ii) MOLECULE TYPE: CDNA (xi) SEQUENCE DESCRIPTION: SEQ ID NO:26: GTATTCCCAA TTNNNNNTAT TTACTACAG 29 INFORMATION FOR SEQ ID NO:27: SEQUENCE CHARACTERISTICS: LENGTH: 29 base pairs o* TYPE: nucleic acid 0 STRANDEDNESS: single TOPOLOGY: linear (ii) MOLECULE TYPE: cDNA (xi) SEQUENCE DESCRIPTION: SEQ ID NO:27: 0 GTATTCCCAA TTNNNNNTAT TTACTACAG 29 138 INFORMATION FOR SEQ ID NO:28: SEQUENCE CHARACTERISTICS: LENGTH: 29 base pairs TYPE: nucleic acid STRANDEDNESS: single TOPOLOGY: linear (ii) MOLECULE TYPE: cDNA (xi) SEQUENCE DESCRIPTION: SEQ ID NO:28: GTAAGTTTAC TANNNNNTTT TCTCCTCAG 29 INFORMATION FOR SEQ ID NO:29: SEQUENCE CHARACTERISTICS: LENGTH: 29 base pairs TYPE: nucleic acid STRANDEDNESS: single TOPOLOGY: linear (ii) MOLECULE TYPE: cDNA (xi) SEQUENCE DESCRIPTION: SEQ ID NO:29: GTAAAAATTA TANNNNNTTT CTTTTTCAG 29 S* INFORMATION FOR SEQ ID SEQUENCE CHARACTERISTICS: LENGTH: 29 base pairs TYPE: nucleic acid 3 STRANDEDNESS: single TOPOLOGY: linear (ii) MOLECULE TYPE: cDNA (xi) SEQUENCE DESCRIPTION: SEQ ID 3 GTATTGTACT TGNNNNNTAT CCTTTGTAG 29 139 INFORMATION FOR SEQ ID NO:31: SEQUENCE CHARACTERISTICS: LENGTH: 29 base pairs TYPE: nucleic acid STRANDEDNESS: single TOPOLOGY: linear (ii) MOLECULE TYPE: cDNA (xi) SEQUENCE DESCRIPTION: SEQ ID NO:31: GTTGCTTTTT CANNNNNTTA TCTAAACAG 29 INFORMATION FOR SEQ ID NO:32: 1 SEQUENCE CHARACTERISTICS: LENGTH: 29 base pairs TYPE: nucleic acid STRANDEDNESS: single TOPOLOGY: linear (ii) MOLECULE TYPE: cDNA (xi) SEQUENCE DESCRIPTION: SEQ ID NO:32: GTACATTTGT CTNNNNNCTT TTCTTTTAG 29 INFORMATION FOR SEQ ID NO:33: SEQUENCE CHARACTERISTICS: LENGTH: 29 base pairs TYPE: nucleic acid 3 STRANDEDNESS: single TOPOLOGY: linear (ii) MOLECULE TYPE: cDNA (xi) SEQUENCE DESCRIPTION: SEQ ID NO:33: 3 GTATCCAGTG TTNNNNNCTT TTTAAACAG 29

Claims (5)

1. An OB receptor protein preparation containing an OB receptor protein, optionally in a pharmaceutically acceptable formulation, said OB receptor protein having part or all of the amino acid sequence according to Seq. ID No. 1 and one or more of the biological properties of naturally occurring OB receptor protein.

2. An OB receptor protein preparation containing an OB receptor protein, optionally in a pharmaceutically acceptable formulation, wherein said OB receptor protein amino acid sequence is selected from among amino acid sequences (a) (b) methionyl residue; (c) methionyl residue; (d) methionyl residue; (e) (f) methionyl residue; (g) methionyl residue; (h) (i) methionyl residue; (j) methionyl residue; (k) methionyl residue; 1-896;

22-896 (according to Seq. ID No. 1): optionally with an N-terminal

23-896 optionally with an N-terminal

29-896 optionally with an N-terminal 1-839; 22-839 optionally with an N-terminal 29-839 optionally with an N-terminal 1-841; 22-841 optionally with an N-terminal 23-841 optionally with an N-terminal 29-841 optionally with an N-terminal 1-891; 141 22-891 optionally with an N-terminal methionyl residue; 23-891 optionally with an N-terminal methionyl residue; 29-891 optionally with an N-terminal methionyl residue; of subparts through further having the C-terminal amino acids, beginning at position 892, of OB receptor B (Seq. ID No. 3) or C (Seq. ID. No. and, a chemically modified derivative of any of subparts through 3. An OB receptor protein preparation of claim 2 wherein said OB receptor protein is further selected from among the OB receptor proteins of subparts through further having the C-terminal amino acids, beginning at position 892, of OB receptor protein D (Seq. ID No. 7). 4. An OB receptor protein preparation of claim 2 wherein said OB receptor protein is further selected from among the OB receptor proteins of subparts through further having substituted the C- terminal amino acids, beginning at position 799, G K F T I L (Seq. ID No. 13). *o*oo *go• 142 An OB receptor protein preparation according to any of claims 1 through 4, wherein the extracellular domain of said OB receptor protein is modified, said modification selected from among: deletion of all or part of the random coil domain; modification of one or both "WSXWS" boxes by substition of the first serine with another amino acid; modification of one or both "WSXWS" boxes by substitution of the last serine with another amino acid; and modification of one or both "WSXWS" boxes by substitution of the first tryptophan with another amino acid. 6. A DNA molecule encoding an OB receptor protein according to any of claims 1-5 selected from the group consisting of: the DNA sequences set forth in Seq. ID nos. 2, 4, 6, 8, 11, 12, and 14; a DNA which selectively hybridizes to a DNA of subpart and 25 a DNA which, but for the degeneracy of the genetic code would hybridize to a DNA of subpart or 7. A biologically functional viral or plasmid vector containing a DNA of claim 6. 8. A procaryotic or eucaryotic host cell containing the vector of claim 7. 9. A host cell modified so that expression of endogenous OB receptor protein is enhanced. 143 A host cell of claim 9 which is an isolated human host cell. 11. A process for producing an OB receptor protein comprised of culturing, under suitable conditions, a host cell according to any of claims 8, 9 or 10, obtaining the OB receptor produced, and optionally preparing a pharmaceutical composition containing said OB receptor. 12. A method of treating an individual for a therapeutic disorder selected from among obesity, diabetes, high blood lipid levels, and high cholesterol levels comprised of administering a therapeutic amount of an OB receptor protein preparation containing an OB o o receptor protein according to any of claims 1-5, or produced by the process according to claim 11. 20 13. A method of treating an individual for oolot weight loss or weight maintenance for solely cosmetic purposes comprised of administering an effective amount *of an OB receptor preparation containing an OB receptor protein according to any of claims 1-5, or produced by the process according to claim 11. 14. Use of an OB receptor protein according to claims 1-5, or produced by the process of claim 11, for manufacturing a medicament for the treatment of obesity, diabetes, high blood lipid levels, or high cholesterol levels. 144 An OB protein/OB receptor protein complex preparation, containing an OB protein moiety and an OB receptor protein moiety, optionally in a pharmaceuti- cally acceptable formulation, wherein: said OB receptor protein is selected from among those set forth in any of claims 1 and 2; said OB protein moiety is selected from among: a naturally ocurring OB protein; and, (ii) a non-naturally ocurring OB protein, analog or derivative thereof. 16. An OB protein/OB receptor protein complex preparation of claim 15 wherein said OB receptor protein is selected from among those set forth in any of claims 3, 4, and 17. A method of treating an individual for a therapeutic disorder selected from among obesity, diabetes, high blood lipid levels, and high cholesterol levels comprised of administering a therapeutic amount 25 of an OB protein/OB receptor protein complex preparation of claims 15 or 16. 18. A method of claim 17 wherein said OB protein/OB receptor protein complex preparation is formed in vivo by administering, into a patient, a first population of cells expressing an OB protein, and a second population of cells expressing an OB receptor protein. 145 19. A method of treating an individual for weight loss or weight maintenance for solely cosmetic purposes comprised of administering a therapeutic amount of an OB protein/OB receptor protein complex preparation containing an OB receptor protein moiety according to any of claims 1-5, or produced by the process according to claim 11. 20. Use of an OB protein/OB receptor protein complex preparation, according to claims 15 or 16, for manufacturing a medicament for the treatment of obesity, diabetes, high blood lipid levels, or high cholesterol levels. Dated this seventh day of February 2001. *S Amgen Inc. Wray Associates Perth, Western Australia Patent Attorneys for the Applicant