AU2004201574B2 - 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 Level 4, The Quadrant 1 William Street Perth, Western Australia Australia Attorney code: WR Invention Title: "OB Protein Receptor and Related Compositions and Methods" This application is a Divisional Application by virtue of Section 79B of Australian Patent Application 18344/01 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 372: 425-432 (1994); see also, 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 things, weight loss. See Generally, Barinaga, "Obese" Protein Slims Mice, Science 269: 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 269: 540-543 (1995); Halaas et al., Science 21d: 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 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. See Stephens et al., supra.

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 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: HI H2 TM Box 1 Box2 Box3

IC

Using the information.provided by Bazan, sura, 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 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 type 3 repeats each, one set of paired cysteine residues each (thought to form a disulfide bridge), and one "WSXWS box'.' (referring to the 6 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 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 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 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 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., Cell j4: 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 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 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, 9 which 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 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 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 Saccharomvces cerevisiae), insect, or procaryote Z. 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 cDNA and genomic genes may be readily accomplished by well-known site-directed mutagenesis techniques and employed to generate analogs and derivatives of OB 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 generally, 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 facilitates purification, such as a poly-histidine tract, an antigenic epitope or a binding domain, may also be present. See, in general Ford et al., Protein Expression and Purification 2: 95-107, 1991, which is herein incorporated by reference.

12 Table 1 Conservative Amino Acid Substitutions Basic: arginine lysine histidine Acidic: glutamic acid asPartic acid Polar: glutamine asoaraaine Hydrophobic: leucine isoleucine valine Aromatic: phenylalanine tryptophan tvrosine Small: glycine alanine serine threonine methionine 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, 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, 78: 3403-3407 (1891); Walter et al., PNAS-USA 78: 4882-4886 (1891); Wong et al., PNAS-USA 21: 5322-5326 (1982); Baron et al., Cell 28: 395-404 (1982); Dressman et al., Nature 295: 185-160 (1982); and Lerner, Scientific American 248: 66-74 (1983). See, 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 OB 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. 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 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 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; 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 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; 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 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: 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 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, 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 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 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 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 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.

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 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 selected from among: (using the numbering of subpart (a) above): amino acids 98-146 (ii) amino acids 1-32 (iii) amino acids 1-35 (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 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; 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 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 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 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 copolymer, 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 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 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. 21: 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 other chemical moiety) may be bound. The amino acid residues having a free amino group may include lysine residues 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).

Preferred 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 N-terminally pegylate the protein by performing the reactionat 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 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 compositions include diluents of various buffer content Tris-HC1, 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 bulking substances lactose, mannitol); incorporation of the material into particulate preparations of polymeric compounds such as polylactic acid, polyglycolic acid, etc. or into liposomes. See, _La, PCT W096/29989, 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, eg., 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 BI12 and Proteins," issued 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 36cyclodextran. Delivery via transport across other mucous membranes is also contemplated.

Dosages 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 or serum) may first be used to determine endogenous levels of OB protein (or receptor). Such diagnostic 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 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 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 supra) 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 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 gall stones, such as ultrasonic or laser methods, may 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 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 Comoositions and Methods As indicated supra, polypeptide products of the invention may be "labeled" by association with a detectable marker substance radiolabeled with 40 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 OB receptor. The 41 hybridoma technique described originally by Kohler and Milstein Eur. J. Immunol. i, 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, (s 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 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 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 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.

Sequencina 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 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 1 0 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 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 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 N0VP1 01; paeQ-= d kl Northern BIot a 4 Kb 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 EXAMPLE 3: IDENTIFICATION OF HUMAN OB RECEPTOR GENOMIC DNA AND CHROMOSOME LOCALIZATION; IDENTIFICATION OF HUMAN OB RECEPTOR "D" The full length human OB receptor genomic DNA was also prepared. OB receptor 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 present form OB receptor protein, the first amino acid of the mature protein is likely (using hydrophobicity 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 841 The transmembrane domain appears to begin at position 840 or 842 The end of the transmembrane domain appears to be located at position 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 83: 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, se 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 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.

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 FTI 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 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 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 38: 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-ut 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.

2 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 phosphorylatont a o signalling molecules.

Methods 1. OB receptor Umolcules. 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. etection of hoshorvlation. 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 37Oc, 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 4GI0, UBI, Lake Placid, NY), and separated by SDS polyacrylamide gel electrophoresis. After electrophoresis and electroblotting to membranes the 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. Result. 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 JAK1 JAK2 JAK3 TYK2 ERKs 1 2 t Antibody detection target 293 cells expressing receptor form treated with 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.

53 EAMPLE 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 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 Eca RI linker and a Bgl II linker) were used to facilitate subcloning.

Use of soluble recombinant human OB receptor PCDA. 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 Bgj 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. 172: 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. £2:1120-1124 (1988). Transient supernatants were harvested from electroporated populations and used to infect tunicamycin treated parental GP+E-86 cells.

Tunicamycin treatment relieves the block to superinfection of the parental packaging cells. G418 (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 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 Suernatants 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 Transolantation 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.

Bone marrow cells were harvested from femurs 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), 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-11), and 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 106 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 (lmg/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.

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), 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 analysis, and weekly thereafter. Food consumption was measured daily on selected groups of individually-housed animals.

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 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 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) da 28 da 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 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.

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 of this example). The data reflect nanograms of OB protein per milliliter of serum, plus or minus the standard error of the mean.

59 TABLE Treatment ExDeriment ExDeriment #2tt OB alone* 2.93 0.77 9.74 1.02 Receptor** 0.08 0.05 0.12 0.07 alone

O

B 12.11 1.90 15.18 2.52 Rece tor*** 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, with OB protein serum levels measured after 38 days.

tt 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 acids 91-100 was found to slightly bind to recombinant human OB receptor protein. The polyclonal antibody prepared against amino acids 874-885 was found not to bind to recombinant human OB receptor protein.

An additional study was performed which 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.

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.

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 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. Ie Winter et al., Nature 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.

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.

Throughout the specification, unless the context requires otherwise, the word "comprise" or variations such as "comprises" or "comprising", will be understood to imply the inclusion of a stated integer or group of integers but not the exclusion of any other integer or group of integers.

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 Rcpo A mn cdS~ec Sa DN.1(m Acid. .ingl etr brvain 1 MICQKFCVVL

LHWEFIYV:T

51 101 L 0 151 201 251 L 5 301 351 ~0 401 451 501 551 601 651 701 751 801 851 901 AGL SKNT SNE LCADNIEGKr

LFRNYNYKVH

PTAKLNDTLL

GNLKISWSSp GS SYE VQVRG

HCIYKKENKI

PRGKFTYDAV

TSTIQSLAES

QP IFLLSGYT

IGLLKISWEK

P DLCAVYAVQ GD TMKKEKNV

FTFLWTEQAH

SSCVIVSWIL

FIPIEKYQFS

SSSILLLGTL

M1ITTDEPNVP

NGHYETAVEP

F VS TVN SLVF LLYVLP EVLE

MCLKITSGGV

PLVPFPLQYQ

KRLDGPG IWS VP SKE IVWWM

YCCNEH-ECHH-

TLQLRYHiRS

S

MWIRINH-SLG

AFNLSYP

IT?

KFNSSGTHFS

QOIDANWNIQ

DSP LV? QKG S

IFQSPLMSVQ

VKYSENSTTV

DWSTPRVFTT

NLAEK IP QSQ RYAELYVI DV

LYCSDIPSIH

SLDSPPTCVL

WRFKLSCPF

NLSKTTFHCC

CWLKGD LKLF

FQMVH-CNCSV

P INNVKP DP P

IREADKIVSA

QDVIYFPPKI

YDVVS DHVS K

NINISCETDG

PISEPKDCYL

PDSVVKPLPP

VQWKMYEVYD

TVVMDIKVPM

INHHTSCNGT l

F'SWPMSKVNI

3EIKWLRIsS )DIEK.HQSDA

C

SWAQGLNFQ

F

KKIQLNF*E L NS TYD YFLLP

FRSEQDRNCS

ICYVESLFKN

HECCECLVPV

LGLHMITDD

TSLLVDS

ILP

L TSVG SNVS F VTFFNLNE

TK

YLTKMTCRWS

QSDGFYECIF

S SVKAE IT IN

.KSKSVSLPV

RGPEFWRI IN

'JSEDVGNH-TK

TQSLSAYPLN

,VKKYYIHDH

~LYVI VPVI I

~RTDIL*SLI

ITYGGLC

*FR

PVFPENNLQF

QIRYGLSGKE

VRCKRLDGLG YWSNWSNPAy TLLWKP LMKN D SLC SVQRYV TVTVLAINS I GASVANFNLT SPSDYKLMYF

IIEWKNLNED

LYPIFMEGVG

KPKIINSFTQ

LISHQRMKKL

FWEDVPNPKN

TSQQSIEY*K IFTF*RRGAN 951 T*NRCVNLGS KCRFESSLDV

*L

65 Human OB Receipr DNA Seauence (Sea, 1D No. 2 (DNA) 1 CCGCCGCCAT CTCTGCCTTC GGTCGAGTTG GACCCCCGGA

TCAAGGTGTA

51 CTTCTCTGAA GTAAGATGAT TTGTCAAAAA TTCTGTGTGG

TTTTGTTACA

101 TTOGGAATTT ATTTATGTGA TAACTGCGTT TAACTTGTCA

TATCCAATTA

151 CTCCTTGGAG

ATTTAAGTT

201 251 301 351 401 451 501 551 601 651 701 751 801 851 901 951 1001 1053.

1101 1151 1201 1251

TACTTCCTT'

ACAT TATGA(

TTTCTAACT'.

GATAGAAAC'

TTCAACAGtl'

TACAGTGCTC

TCAT TATTTP

ATATGTTCTG

GCAGTTTTCA

TGTCTTGTGC

TTTGAAAATC

TTCAGCCCAT

GAAATCACAG

GGTACCATTT

CAGT TATCAG

GACAGTATAC

ACTGGATGGC

CCACACAAGA

TCTAATGTTT

CTCAAAAGAG

GCCAGTATGA

CTGAATGAAA

T TGCCTGCTG(

ACAGCTGTTC

r ATCCAAAACI C GCTCCTTATC

SAATTCTTTAC

GCTAAAAGGP

LAGAATCTATT

CCTGAAGTGT

GATGGTTCAC

CTGTGCCAAC

ACATCTGGTG

AAATATGGTG

ATGATGGTAA

CCACTTCAAT

AGAAGCTGAC

TTCCTGGGTC

CCAGGAATCT

TGTCATATAC

GTCTTGCATGC

3 ACTCTCAAAG .3 AACCTAAGTT k ACTTTCCACT

TGCAGACAAC

TTTTTCAACA

GACTTAAAAT

CAGGAATTAT

TAG.AAGATTC

TGCAATTGCA

AGCCAAACTC

GAGTAATTTT

AAGCCTGATC

TTTAAAGATT

ATCAAGTGAA

AAGATTGTCT

TTCGTATGAG

C

GGAGTGACTC

TTTCCACCTA

CACCAAATTC

A.ATACTTCA-A

TAATTCAAGT

GTTGCTTTCG

ATT GAACGAA

AATAGATGCA

TAT TCATCTG

AACTATAAGG

ACCTCTGGTT

GTGTTCATGA

AACGACACTC

CCAGTCACCT

CACCATTAGG

rCTTGGTCcAC kTATTCAGAG :AGCTACATC

C

TTCAGGTGA

G

;AGTACTCCT

C

LAATTCTGACA

AACCTATGAC

ATTC GAATGG

GGTACTCACT

GAGTGAGCA

AGACATTTGT

AACTGGAACA

TTATGTGGAG

TCCATCTTTT

CCC CAAAAAG ATGT TGTGAA

TCCTTATGTG

:TAATGTCAG

CTTGCATATG

CCCACCAT

T

.aTTCTACAA

CTGCTAGTA

;GGGCAAGAG

:GTGTCTTTA

AGTGTTGGG

CTTTTCACTG

CATCTATAAG

ATTGTTTGGT

GGATGAATTT

TGTTGTGAGT

GATCATGTTA

CCAAACCTCG AGGAALAGTTT AAGGAAAACA

AGATTGTTCC

AGCTGAGAAA

ATTCCTCAAA

GCAAAGTTAC

TTTTTTCAAT

ACCTATGATC CAGTGTACTG 66 1301 CTGCAATGAA OATGAATGOO ATOATOOCTA TGOTGAATTA

TATOTGATTG

1352.

1402.

1451 1501 1551 1601 1651 1701 1751 1801 1851 1901 1951 2001 2051 2101 2151 2201 2251 2301 2351 2401 2451 2501 255.1

ATGTCAATAT

ACTTGCAGAT

GCAATTGAGG

TTOATCCCAT

TATGAATGCA

GATTAGGATC

TCCT TOCTGA

GAAATTAOTA

CTTTCCAGAG

AAGAAGTACA

GTCAGTCtCC

CTGTAAGAGG

OCTACAOAGT

TGGAGAATAA

AOTTTGGAAG

C

ATGTGATA

C

GGAA.ATOACA

C

TACGGTTCTG

G

TAACCTTTTC

A

GCTTATCCTT

T

CAGTGATTACA

AAGATGGTGA Aj I'ATATCCATG

A~

:CCAATATTT

A]

:TCAAGATGA

T.P

CAATATC0TCA

GGTCAAOCAG

TO TGAAAO

TG

TAOAATO CAG TATOATAGGA

GCAGCCTTTA

ATOTGAGCO

TTTTOOAGOO

AATCACTCTC

TTCTGTGGTG

TAAACATTGG

AATAACCTTO

ATGGAAGATG

CAGTTCCAGA

CTAGATGGAC

CGTCATGGAT

~TAATGGAGA

CCCTGATGA

:CATCATACT

GAAATTCAC 91 CCATCAATT

C

TGGCCTATG

A

A.AACAGCAG

T

k.GCTAATGT

A

~TAAAATGG

C

E'CATTTTAT

C

'GGAAGGAG

T

LTTGAAAAA C~

AAAGATTGOT

AATOTTCCTA

TAGGTTCAOT

AAGCCACTGC

ATTATTGAAA

AATTCCAGAT

TATGAGGTTT

C TTGTGTGCA

TGGGATATTG

ATAAAAGTTO

TACTATGAAA

k.AAATGACTC

I

CCCTGCAATG

C

~TTCCTGTGG;

.AATTGGTGC I1 LGCAAAGTAA

A

TGTGTGATT

G

.TTTTATTAT

T

TTAGAATCT

C

CCCATTGAG

A

GGGAAAACC k~ kCCAGAGTG

A'.

ATGGGTAOTT

TCAOTTGOGG

CT OTTO TOAT ATT TGCAGAG

TTIATOTGGCT

T GACTO

TCCA

OTCCATOCAG

ATATCTTGGO

TCGCTATGGT

k".TOATOCAALA 3TCTALTGCTG 3AGTAATTGG

,TATGAGAOG

LAGGAGAAA

P

LTTGTGOAGT

G

,AACATGGTC

A

.OAGAGOAAG

C

TCTGTTGOA

A

TATOGTGOA

G

TTTOOTGGA

T.

SAGTGGAAA

K

rTOATCTOT T2 MGACCAGT

TC

AGATAATT

A;

'GOAGOTTT

AT

AAO TAAA-ATGO

AAAOOAOTTT

ATTOCATOTA

TOAT GOT TTT

AOAOAATOTG

OOAAOATGTG

TO TOAAAGOA

AAAAGOOAOT

TTAAOTGOAA

ATCAAAATO

T

kGCAATCCAG CCOTGAAT

TT

~TGTCACTTT

;TTOAGAGAT

.GAAGATGTG

AOATAOTGT

ATTTTAATT

TOAOTOAGT

ACTATCACO

~TCTTAATG

IAGAAG TAT

:AGTCTTTA

~TAGTTTCA

ATGTAATT

67 2602.

2652.

2702.

2751.

2802.

LO0 2852.

2901.

2952.

3001.

3051.

3002 3151

GTGCCAGTAA

ATCACACCAA

AGAATTGTTC

CTTTGAAGTC

CCAACAGTCT

CAAATC TAAA

TTATGTTGAT

TAGATTTGAG

TTTAAAAGTA

TCATAAAACA

A.AATGTCATC

AACTGCAACA

*TTATTTCCTC

AGAATGAAAA

CTGGGCACAA

TAATCATGAT

ATAGAG TATT AAAAAT TCAG

TTAGAACTTA

TCCAGT

TTGG

GTATTCATGA

TTAAGAAAAT

AAATATGTAG

TCTGACAAAT

TTCCATCTT;

AGCTATTTTC

GGACTTAATT

CACTACAGAT

AGAAGATTTT

T TGAACT TCT

AAATAGATGT

ATGTGTGATT

TTTCTGGCTT

TATGGCTGTT

TAGACAATT

T

TGCTTTAAAA

LTTGCCTTGGA;

GC-AAGATGTT

TTCAGAAGAG

GAACCCAATG

TACATTTTGA

GAGAGTTAAC

GTAAATTTGG

AAT TTTCAAA

TTGATTTGCC

GCTGTCATTA

TGTAATTAGG

ATACAATGAT

CATTATTAAT

CCGAACCCCA

AACGGACATT

TGCCAACTTC

AGAAGGGGAG

ATATGGTGGA

GTTCAAAATG

TCATCTAAAG

ATATTOCTOG

CATATCTATT

TGAACTCTAA

TAT

68 Human OB ReCcemtor Amino Aci d Se ienc= N.3 (Amino 1 MICQKFCVVL LHWEFIYVIT AFNLSYPITP WRFKLSCMPp

NSTYDYFLLP

51 AGLSKNTSNS NGHYETAvEP KFNSSGTHFS NLSKTTFHCC

FRSEQDRNCS

101 LCADNIEGKT FVSTVNSLVF QQIDANWNIO CWTrnVTLr 151 LFRNYNYKVH LLYVLPEVLE DSPLVPQKGs 201 PTAKLNDTLL MCLKITSGGV

IFQSPLMSVQ

251 GNLKISWSSP 301 351 401 451 501 551 601 651 701 751 801 851 901.

951 Cs SYEVQVR(

HCIYKKENX.'

PRGKFTYDA\

TSTIQSLE

QPIFLLSGYT

IGLLKISWER

PDLCAVYAVQ

GDTMKKEKNV

FTFLWTEQAH

S SCVIVSWIL Fl? IEKYQFs

SSSILLLGTL

IQHQ *HVVTTF

LFVLVTSSTV

PLVPFPLQYQ

KRLDGPG

IWS

VP SKE IVWWM

YCCNEHECHH

TLQLRYHRsSS

MWIRINHSLG

PVFPENNLQF

VRCKRLDGLG

TLLWKP LMKN VKYS ENS TTV

DWSTPRVFTT

NLAEKIPQSQ

RYAELYVIDV

LYCSDIPS

IH

SLDSPPTCVL

QIRYGLSGKE

YWSNWSNPAY

DSLCSVQRYV

FQMVHCNCSV

P IN1IMVKPDPP

IREADKIVSA

QDVI YFPKI YDVVS

DWVSK

NINISCETDG

PISEPKDCYL

PDSVVKPLpP VQrgKNYEVYD rvvmDiKvpm INHHTSCNGT

HECCECLVPV

LGLHiMEITDD

TSLLVDSILP

LTSVC SNVSF

VTFFNLNETK

YLTKMTCRWS

QSDGFYECIF

SSVKAEITIN

kKSKSVSLPV GCPEFWRI

IN

TVTVLAINSI CASVANFNLT FSWPMSKVNI

VQSLSAYPLN

SPSDYKLMYF IIEWKNLNED GEIKWLRISS

SVKKYYIHDH

LYP IFMEGVG LI SHQRMKKL

FWSLKQFQKI

LT SLRLRVLR KPKIINSFTQ DDIEKHiQSDA FWEDVPNPKN

CSWAQGLNFQ

SVLIHHGKIK

M*CQQLWSL

*PRKAD

PLLNTPR*SA

GLYVIVPVI

I

KKPLSIFLSS

YFQQQILKRV

TLNQV'KLVK

69 Human OB Rece~tor DNA Searuence (Sea. ID N.4 (DNA f 1 CCGCCGCCAT 51 CTTCTCTGAA 101 TTGGGAATTT 151 201 251 301 351 401 451 501 551 601 651 701 751 801 851 901 951 1001 1051 1101 1151 1201

CTCCTTGGAG

TACT TCC TT T ACAT TAT GAG

TTTCTAACTT

GATAGAAACT

TTCAACAGTA

TACAGTGCTG

TCATTATTTA

ATATGTTCTG

GCAGTTTTCA

TGTCTTGTGC

TTTGAAAATC

TTCAGCCCAT

GAAATCACAG

GGTACCATTT

CAGTTATCAG

GACAGTATAC

ACTGGATGGCC

CCACACAAGA

'I

TCTAATGTTT

C

CTCAAAAGAG

A

GCCAGTATGA

T

CTCTGCCTTC

GTAAGATGAT

ATT TAT GTGA

ATTTAAGTTG

TGCCTGCTGG

ACAGCTGTTG

ATCCAAAACA

GCTCCTTATG

AATTCTTTAG

GCTAAAAGGA

AGAATCTATT

CCTGAAGTGT

GATGGTTCAC

CTGTGCCAAC

ACATCTGGTG

AAATATGGTG

ATGATGGTAA

CCACTTCAAT

%GAAGCTGAC

CTCCTGGGTC

~CAGGAATCT

'GTCATATAC

:TTTTCACTG

LTTGTTTGGTC

'GTTGTGAGTC

*GGTCGAGTTG

GA-CCCOGGA

TTGTCAAAAA

TTCTGTGTGG

*TAACTGCGTT

TAACTTGTCA

TCTTGCATGC

CACCAAATTC

ACTCTCAAAG

AATACTTCAA

AACCTAAGTT

TAATTCAAGT

ACTTTCCACT

GTTGCTTTCG

T GCAGACAAC AT TGAAGGAA TTTTTCAACA

AATAGATGCA

GACTTAAAAT

TATTCATCTG

CAGGAAT TAT AAC TATAAGG TAGAAGATTC

ACCTCTGGTT

TGCAATT.GCA

GTGTTCATGA

TCAAGGTGTA

TTTTGTTACA

TATCCAATTA

AACCTATGAC

ATTCGAATGG

GGTACTCACT

GAGTGAGCA-l

AGACATTTGT

AAC TGGAACA

TTATGTGGAG

TCCATCTTTT

CCCCAAAAAG

ATGTTGTGAA

I

r kGCCAAACTC AACGACACTC

TCCTTATGTG

.AGTAATTTT CCAGTCACCT

CTAATGTCAG

kAGCCTGATC CACCATTAGG

TTTGCATATG

ETTAAAGATT TCTTGGTCCA

GCCCACCATT

TCAAGTGAA ATATTCAGAG

AATTCTACAA

LAGATTGTCT CAGCTACATC

CCTGCTAGTA

'TCGTATGAG GTTCAGGTGA

GGGGCAAGAG

GAGTGACTG GAGTACTCCT CGTGTCTTTA TTCCACCTA AAATTCTGAC

AAGTGTTGGG

ATCTATAAG AAGGAAAACA AGATTGTTCC GATGAATTT AGCTGAGAAA

ATTCCTCAAA

;ATCATGTTA GCAAAGTTAC

TTTTTTCAAT

1251 CTGAATGA CCAAACCTCG AGGAAAGTTT ACCTATGATG

CAGTGTACTG

70 1301 1351 1401

OTGOAATGAA

ATGTOAATAT

ACTTGCAGAT

OATG.AATGOO

CAATATOTCA

GGTCAACCAG

ATOATCGCTA

TGTGAAAO

TG

TACAATCCAG

1451 GCAATTGAGG TATOATAGGA

GOAGOCTTTA

1501 TTCATCCCAT ATOTGAGOCC

AAAGATTGCT

1551 1601 1651 1701 1751 1801 1851 1901 1951 2001 2051 2101 2151 2201 2251 2301 2351 2401 2451 2501 2551

TATGAATGCA

GATTAGGATC

TOOT TOOTGA

GAAATTAOTA

OTT TOOAGAG

AAGAAGTACA

GTCAGTCTCO

OTGTAAGAGG

OOTAOAOAGT

TGGAGAATA

AOTTTGGAAG

ATGTGATAAA

GGAAATOAOAC

TAOGGTTOTG

C

TAAOCTTTTO

GOTTATOOTT

TI

OAGTGATTAO

AAGATGGTGA

A

TATATOOATG

A

OOOAATATTT

A'

CTOAAGATGA T T TTTOOAGOO

AATCAOTOTO

TTOTGTGGTG

TAAAOATTGG

AATAACCOTTO

ATGGAAGATG

OAGTTOOAGA

OTAGATGGAO

TGTOATGGAT

TTAATGGAGA

COOCTGATGA

:CATOATAOT

:GAAATTOAO

OOATOAATT

C

LTGGOOTATG

P

AAAOAGOAG

T

AGOTAATGT

A

ATAAAATGG 0 TOATTTTAT 0 TGGAAGGAG Ti ~TTGAAAAA

C~

AATOTTOO

TA

TAGGTTOACT

AAGOOACTC

ATTATTGAAA

AAT TOOAGAT TATGAGG

TTT

CTTGTGTGOA

TGGGATATTG

ATAAAAGTO

TAO TATGAAA .AAATGACTO

I

E'OTGOAATG

C

L'TTOOTGTGG ~AATTGGTGO

'I

LGOAAAGTAA

A

TGTGTGATT

G

TTTTATTAT

T

TTAGAATOT 0 COOATTGAG

A.

GGGAAAAOO

X~

kCOOAGAGTG

A'

TOO TGAATTA

ATGGGTAOTT

TOACT TOO G O TG TTOTOAT

ATTTOOAGAO

TT"ATOTGOO

T

TGAOTCTOOA

O TOOATCOAG

ATATCTTGGO

TOGOTATOGT

kTGATGCAA 2 3TOTATGOTG

IJ

iAOTAATTOO

P

:TATGAGAGG

A

LAGGAGAAAA

A

TTGTOOAGT

G

AAOATGGTO

A

CAGAGCAAG 0 TOTGTTGOA

A

rATOGTGOA

G

TTOOCTGGA

TI

SAGTGGAAA

AP

rTOATOTGT

TA

MGAOOAGT

TC

LAGATAATT

AA

GOAGGTTT

AT

TATO TOATTO-

AAOTAAAATG

AAAOOAOTTT

ATTOOATOTA

TGATGGTTTT

AOAOAATGTO

OOAAOATGTO

TOTGAAAGOA

kAAAGCCAGT

FTAAGTOOAA

k.TCAAAATCT

TOAGGTGOO

LGOAATOOAG

.OOTGAATTT

.TGTOAOTTT

TTCAGAGAT

GAAGATGTG

COATACTGT

~TTTTAATT

E'OAOTOAGT

C.TATCACC

~TCTTAATG

LAGAAGTAT

AG TOTTTA

.TAGTTTOA

ATOTAATT

71 2601 2651 2701 2751 2801 2851 2901 2951 3001 3051 GTGCCAGTA-. TTATTTCCTC

T.TCOATCTTA

ATCAOACCAA AGAATGAAAA

AGCTATTTTG

AGAATTGTTC CTCGGCACAA

GGACTTAATT

ATCTTTTTAT CAAGCATACA

GCATCAGTGA

GAGCCTGAAA CAATTTCAGA

AGATATCAGT

TAAAGATGAG ATGATGCCAA

CAACTGTGGT

ATCTTGAAAA GGGTTCTCTT

TGTTTTAGTG

TTCTCTGACG CTGAGGGTAC

TCAGGTAACC

ACAACCCTTT GTTAAATACG

CCACGCTGAT

AAACTGGTGA

AGA

TTGCTTGG.

GGAAGATGTT

T TCAGAAGAA

CATGTGGTCC

GTTOATACAT

CTCTOTACTT

ACCAGTTCAA

TATGAGGACG

CAGCAACTCT

CATTATTA-AT

OCGA.ACCCCA

ACGTTTGAGC

TCTTCTTTTG

OATGGAAA

TCAACAACAG

CAGTGTTAAC

AAAGCCAGAO

AAACCAAGTG

72 H~uman OB Recent-or lc", Amino Acid SequenaE (Sac. ID No. 5 (pAin Acid): 2. MICQKFCVVL LHWEFIYVIT AFNLSYPITP WRFFLSC2P?

NSTYDYFLLP

52.

102.

152.

201.

251 301 351 401 451 501.

551 602.

652.

701 751 801 851

AGLSKNTSNS

LCADN IEGKT

LFRNYNYKVR

PTAKLNDTLL

GNLKI SWSSP GS SYEVQVRG

HCIYKKENKI

PRGKFTYDAV

TSTIQSLAES

QP IFLLSGYT IGLLKI SWEK NGHYE TAVEP FVS TVNS LVF

LLYVLPEVLE

MCLKITSGGV

PLVPFPLQYQ

KRLDGPGIWS

VPSKEIVWWM

YCCNEHECHH

TLQLRYH.S

S

MWIRINHSLG

?VEP ENNLQF

KFNSSGTHFS

QQIDANWNIQ

DSP LVP QKG S

IFQSPLMSVQ

VK YSENS TTV DWS TP RVF TT

NLAEKIPQSQ

RYAELYVIDV

LYCSDIPSIH

SLDSPPTCVL

QIRYGLSG.KE

YWSNWS NPAY

DSLCSVQRYV

GASVANFNLT

IIEWKNLNED

KPKIINSFTQI

FWEDVPNPKN

NLSKTTFHCC

CWLKGDLKLF

FQMVHCNCSV

P INMVKPDPP

IRFADKIVSA

QDVI YFPPKI

YDWVSDHVSK

NINISCETDG

PISEPKDCYL

PDSVVKPLPP

VQWKMYEVYD

TVVDI

KVPM

INHHTSCNGT

FSWPMSKVNI

GEIKWLRISS

)DIEKHQSDA

-SWAQGLNFQ

FRSEQDRNCS

ICYVESLFKN

HECCECLVPV

LGLHMEITDD

TSLLVDSILP

LTSVGSNVSF

VTFFNLNETK

YLTKMTCRWS

QSDGFYECIF

SSVKAEITIN

AKSKSVSLPV

RGPEFWRI

IN

WSEDVGNHTK

VQSLSAYPLN

SVKKYYIH-DH

3LYVI VPVI I

<Q,,EGSMFVK

PDLCAVYAVQ

VRCKRLDGLG

GD TMKKEKNV TLLWKP LMKN

FTFLWTEQAH.TVTVLAINSI

SSCVIVSWIL

SPSDYKLMYF

Fl? IEKYQFs LYP IFI4EGVG SSSILLLGTL LISHQRD4KKL SHHHSLISST

QGHKHCGRPQ

PSVRNTQE*S

IKKKKKKLEG

901 951 GPLHRKTRDL CSLVYLLTLP

PLLSYDPAKS

73 Human OB Receptor. DNA Secruence Zea D, N. 6 (D NA) 1 51 101 -0 151 201 251 301 351 ?0 401 451 501 551 601.

651 701 751 801 851 901 951 1001 1051 1101 1151

CCCCGCC;

CTTC TCTGP, T TGGGAATT C TCC TTGGA

TACTTCCTT

ACATTATGA

TTTC TAAC T

GATAGAAAC'

TTCAACAGT2

TACAGTGCTC

TOAT TATTt;

ATATGTTCTC

GCAG TTTTCA

TGTCTTGTGC

TTTGAAAATC

TTCAGCCCAT

GAAATCACAG

GGTACCATTT

CAGTTATCAG

GACAGTATAC

ACTGGATGGC

CCACACAAGA

TCTAATGTTT

CTCAAAAGAG

LT CTCTGCCTTC A GTAAGATGAT 'T ATTTATGTGA G ATTTAAGTTG T TGCCTGCTGG G ACAGCTGTTG T ATCCAAAACA r' GCTCCTTATG k AATTCTTTAG

GCTAAAAGGA

LAGAATCTATT

CCTG.AAGTGT

GATGGTTCAC

CTGTGCCAAC2 ACATCTGGTG

C

AAATATGGTG

ATGATGGTAA

T

CCACTTCAAT

A

AGAAGCTGACA

TTCCTGGGTC

T

CCAGGAATCT

G

TGTCATATAC

T

CTTTTCACTG

C~

ATTGTTTGGT G(

GGTCGAGT'

TTGTCAAAI

TAACTGCGI

TCTTGCATC

ACTCTCAAA

AACCTAAGT

ACTTTCCAC

TGCAGACAA

TTTTTCAAC,

GACTTAAAA'

CAGGAATTA'.

TAGAAGATT(

rGCAATTGCI kGCCAAACTC

,AGTAATTTT

LAGCCTGATC

'TTAAAGATT

.TCAAGTGAA

AGATTGTCT

TCGTATGAG

GAGTGACTG

TTCCACCTA

kTCTATAAG 3ATGAATTT

GACCCOGGA

TTCTGTGTGG

TAACTTGTCA

CACCAAATTC

AATAC TTCAA

TAATTCAAGT

GTTGCTTTCG

ATTGAAGGAA

AATAGATGCA

IATTCATCTG

AACTATAAGG

kCCTCTGGTT 3TGTTCATGA2 ACGACACTC

I

:CAGTCACCT

C

:ACCATTAGG

TI

'CTTGGTCCA

G

*TATTCAGAGA

AGCTACATC

C

TTCAGGTGA

G

AGTACTCCT

C

TCAAGGTGTA

TTTTGTTACA

TATCCAATTA

AACC TAT GAO

ATTCGAATOG

GGTACTCACT

GAG TGAGCAA

AGACATTTGT

AACTGGAACA

TTATGTGGAG

ICCATCT TT T

CCCCAAAAAG

kTGT TGTGAA CCT TATGTG

TAATGTCAG

TTGCATATG

;CCCACCATT

ATTCTACAA

CTGCTAGTA

GGGCAAGAG

AAATTCTGAC AAGTGTTGGG

AAGGAAAACA

AGATTGTTCC

1201 GCCAGTATGA TGTTGTGAGT GATCATGTTA GCAAAGTTAC

TTTTTTCAAT

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

CTGAATGAA

CTGCA.ATGk.

ATGTCAATA'.

ACT TGCAGA'

GCAATTGAG(

TTCATCCCA]

TATGAATC

GATTAGGATC

TCCT TCCTGA GAAAT TACTA

CTTTCCAGAG

AAGAAGTACA

GTCAGTCTCC

CTGTAAGAGG

CCTACACAGT

TGGAGAATAA

ACTTTGGAAG

ATGTGATAAA

GGAAATCACA

TACGGTTCTG

TAACCTTTTC

GCTTATCCTT

CAGT GATTAC k~ CCAAACCTC k. CATGAATGC r CAATATCTC.

r' GGTCAACCA, 3TATCATAGG

ATCTGACC

TTTTCCAGC(

AATCACTCT(

TTCTGTGGTC

*TAAACATTGC

AATAACCTTC

AT CGAAGAT C

CAGTTCCAGA

CTAGATGGAC

TGTCATGGAT

TTAATGGAGA

CCCCTGATGA

CCATCATACT

CGAAATTCAC

GCCATCAATT

ATGGCCTATG

TAAACAGCAG

AAGCTAATGT

G AGGAAAGTTT ACCTATGATG

CAGTGTACTC

C ATOATOOCTA TC-CTGAATTA

TATCTGATTG

A TGTGAAACTG ATGGGTACTT

AACTAAAATG

S TACAATCCAC TCACTTCCGG

AAAGCACTTT

k GCAGCCTTTA CTCTTCTGAT

ATTCCATCTA

AAAGATTCCT ATTTCCAGAG

TGATGGTTTT

:AATCTTCCTA TTATCTGGCT

ACACAATGTG

'TAGGTTCACT TGACTCTCCA

CCAACATGTG

AAGCCACTGC CTCCATCCAG

TGTCAAAGCA

ATTATTGAAA ATATCTTGGG

AAAAGCCAGT

AATTCCACAT TOOCTATOCT

TTAAGTGGAA

TATGAGGTTT ATGATGCAAA

ATCAAAATCT

*CTTGTGTGCA GTCTATCCTG

TTCAGGTCCG

TGGCATATTG GAGTAATTGG

AGCAATCCAG

ATAAAAGTTC CTATCAGAGG

ACCTGAATTT

TACTATGAAA AAGGAGAAAA

ATGTCACTTT

AAAATGACTC ATTCTGCAGT

GTTCACAGAT

TCCTGCAATG GAACATGCTC

AGAAGATGTG

TTTCCTGTGG ACAGAGCAAO

CACATACTGT

CAATTGGTGC TTCTGTTGCA

AATTTTAATT

AGCAAAGTAA ATATCGTGCA

GTCACTCAGT

TTGTGTGATT GTTTCCTGGA TACTATCACC ATTTTATTAT TGAGTGGAAA AA.TCTTAATG CTTAGAATCT CTTCATCTGT

TAAGAAGTAT

CCCCATTGAG AAGTACCAGT

TCAGTCTTTA

TGGGAAAACC AAAGATAATT.

AATAGTTTCA

CACCAGAGTG ATGCAGGTTT

ATATGTAATT

AA.GATGGTGA

AA.TAAAATGG

TATATCCATG

ATCATTTTAT

CCCAATATTT

ATGGAAGGAG

CTCAAGATGA

TATTGAAAAA

75 2601 2651 2701 2751 LO0 2801 2851 2901 L 5 2951 GTCCAGTAA TTATTTCCTC TTCCATCTTA TTIGCTTG,

CATTATTZZT

ATCACACCAA ACAATGAAAA AGCTATTTTG GGAAGATGTT

CCGAACCCCA

AGAATTGTTC CTGGGCACAA GGACTTAATT TTCAGAAGAT

GCTTGAAGGC

AGCATGTTCG TTAAGAGTCA TCACCACTCC CTA-ATCTCAA

GTACCCAGGG

ACACAAACAC TGCGGAAGGC CACAGGGTCC TCTGCATAGG

AAAACCACAG

ACCTTTGTTO ACTTGTTTAT CTGCTGACCC TCCCTCCACT

ATTGTCCTAT

GACCCTGCCA AATCCCCCTC TGTGAGAAAC ACCCAAGAAT

GATCAATAAA

AAAAAA AAAAAACTCG

AGGGGG

76 Human OB R-eceotor Amino Acd Seuem Seence 11) No-, 7 1 MICQKFCWVL LEWEFIYVIT A.FNLSYPIT? WRFKLSCNpP

NSTYDYFLLP

51. AGLSKNTSN.

101 LCADNIEGK .0 151 201 251 301 351 ~0 401 451 501 551 601 0 651 701 751 801 851 901 951 LFE NYNYKVI

PTAKLNDTLI

GNLKISWSSE

GSSYEVQVRC

HCIYKKENKI

PRGKFTYDA'V

TSTIQSLAEs

QPIFLLSGYT

IGLLKI SWEK

PDLCAVYAVQ

GDTMKKEKNV

FTFLWTEQAHi S SCVIVSWIL

FIPIEKYQFS

SSSILLLGTL

KHTASVTCGP

GSVCISDQFN

S NGHYETAVEP T FVSTVNSLVF i LLYVLPEVLE

MCLKITSGGV

KRLDGPGIWS

VPSKEIVWWM

YCCNEHECHH

TLQLRYHRS

S

MWIRINHSLG

PVFPENNLQF

VRCKRLDGLG

TLLWKPLMKN

TVTVLAINSI

SPSDYKLMYF

LYPIFMEGVG

LISI-QRMKKL

I

LLLEPETISE

E

SVNFSEAEGT E

KFNSSGTHFS

QQIDANWNIQ

DSPLVPQKGs

IFQSPLMSVQ

VKYS ENS TTV

DWSTPRVFTT

NLAEK IP QSQ

RYAELYVIDV

LYCSDIPSIH

S LD SFPT CVL

QIRYGLSGKE

YWSNWSNPAY

DSLCSVQRYV

GASVANFNLT

IIEWKNLNED

<PKIINSFTQI

'WEDVPNPKN

)ISVDTSWKN

VTYEDESQR

C

DSFSNSSWE

I

FNDKKSIYYL

HFVENNINL

C

LQICVIMGN I NLS KTTFNCC

CWLKGDLKLF

FQMVHCNCSV

PINMVK<PDPP

IREADK

IVSA

QDVIY-FPPKI

YDVVSDHVSK

NINISCETDG

P ISEPKDCyL

PDSVVKPLPP

VQWKMYEVYD

TVVMDIKVPM

INHHTSCNGT

E'SWPMSKVNI

GEIKWLRISS

DDIEKHQSDA

C

:SWAQGLNFQ

P

(DEITTVV

S

~PFVKYATLI

S

:EAQAFFILS

D

rVTSIKXRESG TSSKKTFAS

Y

KCNRL*LWV G

FRSEQDRNCS

IC YVE SLFKN

HECCECLVPV

LGLHil

ITDD

TSLLVDS 112

LTSVGSNVSF

VTFFNLNETK

YLTKIMTCRWS

QSDGFYECIF

35 VKAE IT IN

AKSKSVSLPV

RGPEFWRI

IN

6'SEDVGNHTK

IQSLSAYPLN

3VKK YY IHDH LYVIVPVI

I

PETFEHLF

I

LLS TTDLEK ;NSKP SETGE

QHPNIISPH

.VLLTDKSRV

MPQFQTCST

ERKETRVKF

1001 EQGLINSSVT

KCFSSKNSPL

1051 1101 '0 1151 1201 LTFSEGLDEL

LKLEGNFPEE

SCPFPAPCLF TDIRVLQDsc QTHKIMNKM

CDLTV*FH*R,

ENNCSK*KXK

KKNSRPARPD

77 Hu'man OB Rec-O-tor "Dn'NUCleic Acid. Secrunce (Senr~e ID No.8) 1 GCGGCCGCCA GTGTGATGGA TATCTGCAGA

ATTCGGCTTT

51 101 151 0 201 251.

301 351 401 .0 451 501 551 601 651 701 751 801 851 901 951 1001 1051 1101 1151 1201 1251 GGTCGAGT Ti

TTGTCAAAA

TAACTGCGT'.

TCTTGCATGC

GCTCTCAAA(

AACC TAAGT I

ACTTTCCACI

TGCAGACAAC

TTTTTCAACA

GACT TAAAAT

CAGGAATTAT

TAGAAGATTC

TGCAATTGCA

AGCCAAACTC

GAGTAATTTT

AAGCCTGATC

TTTAAAGATT

ATCAAGTGAA

AAGATTGTCT

TTCGTATGAG

GGAGTGACTG

TTTCCACCTA

CATC TATAAG GACCCCCGGA

TCAAGGTGTA

TTCTGTGTGG

TTTTGTTACA

r TAACTTGTCI

'CACCAAATT(

3 AATACTTCAI

TAATTCAAGI

GTTGCTTTCC

ATTGAAGGA-Z

AATAGATGCA

TATTCATCTG

AACTATAAGG

ACCTCTGGT

T

GTGTTCACGA

AACGACACTC

CCAGTCACCT

CACCATTAGG

TCTTGGTCCA

ATATTCAGAG

CAGCTACATC

kTATCCAATTA

A.ACCTATGAC

k. ATTCGAATGG

GGTACTCACT

GAGTGAGCAA

AGACATTTGT

.AACTGGAACA

TTATGTGGAG

TCCATCTTTT

CCC CAAAAAG

ATGTTGTGAA

TCCTTATGTG

CTAATGTCAG

TTTGCATATG

GCCCACCATT

AATTCTACAA(

CCTGCTAGTAC

C TTC TCTGAA

TTGGIGAATTT

C TCCT TGGAG

TACTTCCTTT

ACATTATGAG

T TTC TAACT T

GATAGAAACT

TTCAACAGTA

TACAGTGCTG

TCATTATTTA

ATATGTTCTG

GCAGTTTTCA

TGTCTTGTGC

rTTGAAAATC2 rTCAGCCCAT 3AAATCACAG

I

3GTACCATTT

C

AGTTATCAG

A

'ACAGTATAC

TI

kCTGGATGGC

C

:CACACAAGA

T

'CTAATGTTT

C

TCAAAAGAG

A

CCAGTATGA Ti TGAATGAAA

C

CTCTGCCTTC

GTAAGATGAT

ATTTATGTGA

ATTTAAGTTG

TGCCTGCTGG

ACAGCTGTTG

ATCCAMAACA

GCTCCTTATG

AATTCTTTAG

GCTAAAAGGA

AGAATCTATT

CCTGAAGTGT

GATGGTTCAC

TGTGCCAAC

kCATCTGGTG LkATATGGTG

TGATGGTAA

~CAC TTCAAT

LGAAGCTGAC

TCCTGGGTC

CAGGAATCT

GTCATATAC

TTT TCACTG

TTGTTTGGT

GTTGTGAGT

CAAACCTCG

GTTCAGGTGA

GGGGCAAGAG

GAGTACTCCT

CGTGTCTTTA

AAATTCTGAC

AAGTGTTGGG

AAGGAAAACA AGATTGTTCC GGATGAATTT AGCTGAGAAA

ATTCCTCAAA

GATCATGTTA GCAAAGTTAC

TTTTTTCAAT

78 1301 1351 1401 1451 LO0 1501 1551 1601 1651 1701 1751 1801 1851 1901 1951 2001 2051 2101 2151 2201 2251 2301 2351 2401 2451 2501

AGGAAAGTTT

ATCATCGCTA

TGTGAAACTG

TACAATCCAG

GCAGCCTTTA

AAAGATTGCT

AATCTTCCTA

TAGGTTCACT

AAGCCACTGC

ATTATTGAAA

AAT T CCAT

TATGAGGTTT

CT TGTGTGCA

TGGGATATTG

ATAAAAGTTC

TACTATGAAA

AAAATGACTC

TCCTGCAATG

TTTCCTGTGG

CAATTGGTGC

AGCAAAGTAA2 TTGTGTGATT ATTTTATTAT

'J

CTTAGAATCT

C

CCCCATTGAG

ACCTATGATG

TGCTGAATTA

ATGGGTACTT

TCACT TGCGG

CTGTTCTGAT

ATTTGCAGAG

TTATCTGGCT

TGACTCTCCA

CTCCATCCAG

ATATCTTGG

TCGCTATGGT

ATGATGCAAA

GTCTATGCTG

GAGTAATTGG

CTATGAGAG

AAGGAGAAA

PTTGTGCAGT

GAACATGGTC2 kCAGAGCAAGC rTCTGTTGCA kTATCGTGCA

C

TTTCCTGGA TI

'GAGTGGAAA

~TTCATCTGT TI LAGTACCAGT T

CAGTGTACTG

TATGTGATTG

AAC TAAAATG

AAAGCACTTT

ATTCCATCTA

TGATGG TTTT ACACAAT GTG

CCAACATGTG

TGTGAAAGCA

AAAAGCCAGT

T TAAGTGGAA

ATCAAAATCT

TTCAGGTGCG

AGCAATCCAG

k~CCTGAATTT kTGTCACTTT 3TTCAGAGAT kGAAGATGTG

~ACATACTGT

~aTTTTAATT ~TCACTCAGT

C

ACTATCACC

C

,ATCTTAATG AAGAAGTAT T CAGTCTTTA C

CTCGCAATGAA

ATGTCAATAT

ACTTGCAGAT

GCAATTGAGG

TTCATCCCAT

TAT GAATGCA

GATTAGGATO

TCC TTCCTGA

GAAATTACTA

CTTTCCAGAG

AAGAAGTACA

GTCAGTCTCC

CTGTAAGAGG

CC TACACAG T

TGGAGAATAA

PkCTTTGGAAG kTGTGATAAA 3GAAATCACA rACGGTTCTG

CAACCTTTTC

CTTATCCTT

:AGTGATTAC2 LAGATGGTGA2 ATATCCATG

I

:CCAATATTT ;Z CAT GAATCC

CAATATCTCA

GGTCAACCAG

TATCATAOGA

ATCTGAGCCC

TTTTCCAGCC

AATCACTCTC

TTCTGTGGTG

TAAACAT TGG

AATAACCTTC

ATGGAAGATG

CAGTTCCAGA

CTAGATGGAC

TGTCATGGAT

T TAATGGAGA

CCCCTGATGA

CCATCATACT

CGAAATTCAC

GCCATCAATT

kTGGCCTATG

CAAACAGCAG

kAGCTAATGT

ATAAAATGG

~TCATTTTAT

TGGAAGGAG

2551 TGGGAAAACC AAAGATAATT AATAGTTTCA CTCAAGATGA

TATTGAAAAA

79 2601 CACCAGAGTG ATGCAGGTTT ATATOTAATT GTGCCAOTAA

TTATTTCCTC

2651 2702.

2752.

28021 28521 2901 2951 3001 3051 32.01 3151 3202.

3251 3301 3351 3401 3451 3501.

3551

TTCCATCTTI

AGCTATTTTC

GGACTTAATI

TACAGCAT.P

CAGAAGATAT

CCAACAACTO

TGTTTGTATT

GTACTGAGGT

TACGCCAcC

AGGGCTTATA

COTTGAAGGA

TTTTTTATAT

ATTCTCAGAA

AAGAAAATAA

AAAAAGAGAG

CCCATTCCCA

GTTGCTCACA

AAGACTTTTO

TCATAAGATC

~TTGCTTGA;

OOAAOATGTI

TTCAOAAGCC

GTGACATGTG

CAGTOTTGAT

TGGTCTCTCT

AGTOACCAGT

AACCTATGAG

TOATCAGCAA

AATAGTTCAG

TTCTTTCTCT

TATCOGATCA

GOAT TGGATG

TGATAAAAAG

AGAGTGGTGT

GCCCCCTGTT

CATTATTAAN

*CCGAACCCC;

*AGAA-AOGTT1

GTCCTCTTCI

ACAT CAT GO.

ACTTTCAACA

TCAACAGTOT

GACGAA-AOCC

CTCTAAACCA

TCACCAAGTG

AATAGCTCAT

OCATCCCAAC

AACTTTTGAA

TCTATCTATT

GCTTTTGACT

TATTCACGGA

ATCACACCAA.

AGAATGAAAA

AGAATTGTTC

CTGGOCACAA

GAGCATCTTT

TTATCAAOCA

TTTOGAOCCT

OAAACAATTT

AAA-ATAAAOA

TOAGATOATO

ACAOATCTTO AAAAGGTTO

TAACTTCTCT

AGAOACAACC

AG TGAAAC TO CTT CTO TAGC

OOOAOATAOA

ATAATTTCAC

ATTOOAOOOA

ATTTAOOOOT

OACAAOTCAA

CAT CAGAGT T

ACTTAOOAAC

CAAACTTOTT

CCTAACTGTG

GAGOCTGAGG

OTT T TTAAA

OTGAAGAACA

AAAA-ATTCTC

GOCCCAOOCA

CACACCTOAC

AATTTCCCTO

CACCTCAATC

GOTATOOTO

OTCCAGOACA

TTCTAGTAAO

C TAO TCAGAC

TAATCTAGA

CTTTOTAOAA

AATAATATCA

CATCTTACAT

OCCTCAATTC

ATOOAAAACA AOATGTGTGA 80 Human OB RecetorProtein mOSOmal DN(e Intron 1 Intron 2 tacctttccag OAT TGG G His Trp Glu 12 13 14 gtaagttatttg a..tacctaa-ag

AA

CAA ATh Gin Ile 122 123 G gtaagcactagc Intron 3 ttttaaattcag TAT GTT Tyr Val 163 164 CT gtaaqtaccaaa Leu Intron 4 ttttcaatatag AAT ATG G gtaagttatgca.....

Asn Met Val 233 234 235 Intron 5 tttttccttaag GTG TAO Tc TTT ATT Phe Ile 16 AT GCA AAC Ala Asn 125 126 G COT GAA Pro Glu 166 167 TG AAG OCT Lys Pro 236 237 GCT GAC AAG Ala Asp Lys 284 285 286 AT GTC ATA Val Ile 333 334 AT GIC AAT Val Asr 430 431 G AGO AGO Ser Ser 469 470 TG AAG CCA Lys Pro 536 537 ATC AGA Ile Arg 281 282 ACA CAA Thr Gin 330 331 GAA gtaagtatattt.....

Glu 283 G gtaggttatgta.....

Asp Intron 6 aatatttaacag Iniron 7 ccctcattacag GTG ATT G gtaagaaaacag Val Ile Asp 427 428 429 TAT CAT AG gtactattati.....

Tyr His Arg 466 467 468 TCT GTG G gtatgtcaagct Ser Val Va.

533 534 535 CAA TGG AAG gtaccttttact...

Gin Trp Lys 582 583 584 ATA AAA G gtctgcagagat Ile Lys Val 636 637 638 Intron 8 tgtttcaaatag Intron 9 tatctttaaag Intron 10 aaaaatttctag Intron 11 cttatttt aca g ATG TAT GAG Met Tyr Glu 585 586 587 TT CCT ATG Pro Met 639 640 Intron 12 gtcattttgqag 81 CTT TGG AAG gtattcccaatt Leu Trp Lys 663 664 665 AGC AAA G gtaagaagaggt..

Ser Lys Val 0 736 737 738 ATC CAT G gtaagtttacta Ile His Asp 797 798 799 ACT CAA G gtaaaaattata Thzr Gln Asp .0 829 830 831 CAC CAA AG gtattgtacttg His Gin Arg ~5 864 865 866 TTT CAG AAC gttgctttttca Phe Gin Lys ~0 889 890 891.

Exon A AAA TAT GAT grtacatttgtct Intron 13 tatttactacag Intron 14 ttcccctcag Intron 15 ttttctcctcag Intron 16 ttctttttcag Intron 17 tatcctttgtag CCC CTG ATr, Pro Leu Met 666 667 668 TA AAT ATC As n Ile 739 740 AT CAT Trrr His Phe 800 801 AT CAT ATT Asp Ile 832 833 A ATG AAA Met Lys 867 868 Fxon A AGA ACG CAC Arg Thr Asp 892 893 894 E-xon D CCA GAA ACG Pro Giu Th~r 892 893 894 Exon B AAA CT TTC Lys Arg Leu 892 893 894 E~xon C ATG CTT GAA Met Leu Giu 892 893 894 Intron 18 ttatctaaacag Intron 18 cttttcttttag Exon D GAA ACC AGA gtatccagtgtt...

Iniron 18 ctttttaaacag 82 Human 05 Receptor Protein, Recombinan Sgecreted Receator amino acid secgience (Sea. ID. No. 1 MICQKFCVVL

LHWEFIYVIT

AFNLSYPITP WRFKLSCNMP

NSTYDYFLLP

51 101 151 201 251 301 351 401 451 501 551 601 651 701 751 AGLS KNT SN S LCADNIECa'KT

LFRNYNYKVH

P TAKLNDTLL

GNLKISWSSP

GSSYEVQVRG

HCIYKKENKI

PRGKFTYDAV

TSTIQSLAES

QP IFLLSGYT

IGLLKISWEK

P DLCAVYAVQ GD TMKKEKNV FTFLWTEQAiH S SCVI VSWI L

NGHYETAVEF

FVSTVNSLVF

LLYVLP EVLE

MCLKITSGGV

PLVPFPLQYQ

KRLDGP GIWS

VPSKEIVWWM

YCCNEHECHH

TLQLRYHRS S

MWIRINHSLG

PVFPENNLQF

VRCKRLDGLG

TLLWKP LMKN

TVTVLAINSI

SPSDYKLMYF

KFNSSGTNFS NLSKTTFHCO

FRSZQDRNCS

QQIDANWNIQ CWLKGDLKLF

IOYVESLFKN

DSPLVPQKGS FQMVHCNCSv

H-ECCECLVPV

IFQSPLMSVQ PINMVKPDPP

LGLHMEITDD

VKYSENSTTV IREADKIVSA

TSLLVDSILP

DWSTPRVFTT QDVIYFPPKI

LTSVGSNVSF

NLAEKTPQSQ YDVVSDHVSK

VTFFNLNETK

RYAELYVIDV NTNISCETDG

YLTKMTCRWS

LYCSDIPSIH PISEPKDCyL

QSDGFYECIF

SLDSPPTCVL PDSVVKPLpP

SSVKAEITIN

QIRYGLSGKE VQWKMYEVYD

AKSKSVSLPV

YWSNWSN4PAY TVVNDIKVPM

RGPEFWRIIN

DSLCSVQRYV INHHTSCNGT

WSEDVGNH-TK

GASVANFNiLT FSWPMSKVNI

VQSLSAYPLN

IIEWKNLNED GEIKWLRISS SVKKYYIHDH- 801 FIPIEKYQFS LYPIFMEGVG KPKIINSFTQ

DDIEKHQSD

83 Human-OB Receor- roin eobnn-ertdRcoo

N

secmence (Seq. ID. No. 11):.

2. GCOGCCGCCA GTGTGATGGA TATCTGCAGAk ATTCGGCTTT

CTCTGCOTTC

52. GGTCGAGTTG 101 TTGTCAAAAA

GACCCCCGGA

TTCTGTGTGG

TCAAGGTGT

TTTTGTTAC1 151 201 251.

301 351 0 401 451 501 551 601 651 701 751 801 851 901 951 1001 1051 1101 1151 1201

TAACTGCGT

TCTTGCATG(

GC TC TCAAAC

AACCTAAGTI

ACTTTCCACT

TGCAGACAAC

TTTT TCAACA CACT TAAAAT

CAGGAATTAT

TAGAAGATTC

TGCAATTGCA

AGCCAAACTC

GAGTAATTTT

AAGCCTGATC

TTTAAAGATT

ATCAAGTGAA

AAGATTGTCT

TTCGTATGAG

GGAGTGACTG

C TAACTTGTCA

TATCCAATTI

CACCAAATTC

AACCTATGAC

AATACTTCAA

ATTCGAATGC

TAATTCAAGT

GGTACTOACT

GTTGCTTTCG

GAGTGAGCAA

ATTGAAGGAA

AGACATTTGT

AATAGATGCA

AACTGGAACA

TATTCATCTG

TTATGTGGAG

AACTATAAGG TCCATCTTTT ACCTCTGGTT

CCCCAAAAAG

GTGTTCACGA

ATGTTGTGAA

AACGACACTC

TCCTTATGTG

CCAGTCACCT

CTAATGTCAG

CACCATTAGG

TTTGCATATG

TCTTGGTCCA

GCCCACCATT

ATATTCAGAG

AATTCTACAA

CTTCTCTGAA

GTAAGATGAT

k TTGGGAATTT

ATTTATGTGA

k CTCCTTGGAG

ATTTAAGTTG

:TACTTCCTTT

TGCCTGCTGG

;ACATTATGAG

ACAGCTGTTC

TTTCTAACTT

ATCCAAAACA

GATAGAAACT

GCTCCTTATG

TTCAACAGTA

AATTCTTTAG

TACAGTGCTG

GCTAAAAGGA

TCATTATTTA

AGAATCTATT

ATATGTTCTG

CCTGAAGTGT

GCAGTTTTCA

GATGGTTCAC

TGTCTTGTGC

CTGTGCCAAC

TTTGAAAATC

ACATCTGGTG

TTCAGCCCAT

AAATATGGTG

GAAATCACAG

ATGATGGTAA

GGTACCATTT

CCACTTCAAT

CAGTTATCAG AGAAGCTGAC CAGCTACATC CCTGCTAGTA

GACAGTATAC

GTTCAGGTGA GGGCCAAGAG

ACTGGATGGC

GAGTACTCCT CGTGTCTTTA

CCACACAAGA

TTCCTGGGTC

CCAGGAATCT

TGTCATATAC

CTTT TCACTG

ATTGTTTGGT

TOT TGTGAGT TTTCCACCTA

AAATTCTGAC

CATC TATAAG AAGGAAAACA GGATGAATTT

AGCTGAGAAA

AAGTGTTGGG

TCTAATGTTT

AGATTGTTCC

CTCAAAAGAG

ATTCCTCAAA GCCAGTATGA 84 1251.

1301 1351 1401 .0 1451 1501 1551 1602.

1651 ?0 1701 1751 1801 1851 1901 1951 2001 2051 2101 2151 2201 2251 2301 GAT CATGTTA

AGGAAAGTTT

ATCATCGCTA

TGTGAAACTG

TACAATCCAG

GCAGCCTTTA

AAAGATTGCT

AATCTTCCTA

TAGGTTCACT

AAGCCACTGC

ATTATTGAAA

AATTCCAGAT

TATGAGGTTT

CTTGTGTGCA

TGGGATATTG

ATAAAAGTTC

TACTATGAAA

AAAATGACTC

TCCTGCAATG

TTTCCTGTGG

CAATTGGTGC

AGCAAAGTAA

GCAAAGTTAC

ACCTATGATG

TGCTGAATTA

ATGGGTACTT

TCACTTGCGG

CTGTTCTGAT

ATTTGCAGAG

TTATCTGGCT

TGACTCTCCA

CTCCATCCAG

ATATCTTGGG

TCGCTATGGT

ATGATGCAAA

GTCTATGCTG

GAGTAATTGG

CTATGAGAGG

AAGGAGAAA

ATTGTGCAGT

GAACATGGTC

ACAGAGCAAG

TTCTGTTGCA

ATATCG TGCA

TTTTTTCAAT

CAGTGTACT-

TATGTGATTG

AACTAAAATG

AAAGCACTTT

ATTCCATCTA

TGATGGTTTT

ACACAATGTG

CCAACATGTG

TGTGAAAGCA

AAAAGCCAGT

TTAAGTGGAA

ATCAAAATCT

TTCAGGTGCG

AGCAATCCAG

ACCTGAATTT

ATGTCACTTT

GTTCAGAGAT

AGAAGATGTG

CACATACTGT

AA.TTTTAATT

GTCACTCAGT

TACTATCACC

CTG.AATGAAA COAAACCTC CTGCAATGAA CATGAATGCC ATGTCAATAT CAATATCTCA ACT TGCAGAT GGTCAACCAG GCAATTGAGG

TATOATAGGA

TTCATCCCAT ATOTOACCC TATGAATGCA TTTTCCAGCC GATTAGGATC AATCACTCTC TCCTTCCTGA TTCTGTGGTG GAAATTACTA TAAACATTGG CTTTCCAGAG AATA.ACCTTC AAGAAGTACA ATGGAAGATG GTCAGTCTCC CAGTTCCAGA

CTGTAAGAGG

COTACACACT

TGGAGAATAA

ACTTTGGAAG

ATGTGATAAA

GGAAATCACA

TACGGTTCTG

TAACCTTTTC

GCTTATCCTT

CAGTGATTAC

AAGATGGTGA

CTAGATGGAC

TGTCATGGAT

T TAATGGAGA

CCCCTGATGA

CCATCATACT

CGAAATTCAC

GCCATCAATT

ATGGCCTATG

TAAACAGCAG

AAGC TAATGT

AATAAAATGG

2351. TTGTGTGATT GTTTCCTGGA 2402.

2451 2501 ATTTTATTAT TGAGTGGAAA AATCTTAATG CTTAGAATCT CTTCATCTGT TAAGAAGTAT CCCCATTGAG AAGTACCAGT TCAGTCTTTA TATATCCATG ATCATTTTAT CCCAATATTT ATGGAAGGAG 85 2551 TGGGAAAACC AAAGATAATT AATACTTTCA CTLCAAGATGA

TATTCAAAAA

2601 CACCAGAGTG ATTGATAAGG

ATCC

86 Human OB Rercentor Protein. Recombinant Secred Rcepntor DNA secuence with C-t-prrninpl FLAG (SeaT. Tn. No. 1?2) 1 51 101 151 201 251 301 351 401 451 501 551 601 651 701 751 801 851 901 951 1001 1051 1101 1151 COAT TGAAGT

CCAAAATGTC

TGTACGGTGG

CAATGGGAGT

GTAATAACCC

GAGGTCTATA

TTOTTTTGGC ACCAAAATCA

ACGGGGATTT

CGCCCCGTTG ACCAAATGG

GCGGTAGGCG

TAAGCAGAGC TCGTTTAGTG AACCGTCAGA TCTCTAGAAG CTGGGTACCA GCTGCTAGCA AGOTTOCTAC COGCOCCAG TGTGATGGAT ATCTGCAGAA TTCGGCTTTC TCTGCCTTCG GTCGAGTTGG ACCCGGAT CAAGGTGTAC TTCTCTGAAG TAAGATGATT TGTCAAAAAT TCTGTGTGGT TTTGTTACAT TGGGAATTTA TTTATGTCAT AACTGCGTTT AACTTGTCAT ATCCAATTAC TCCTTGGAGA TTTA.AGTTGT CTTGCATGCC ACCAAATTCA ACCTATGACT ACTTCCTTTT GCCTGCTGGG CTCTCAAAGA ATACTTCAAA TTCGAATGGA CATTATGAGA CAGCTGTTGA ACCTA.AGTTT AATTCAAGTG GTACTCACTT TTCTAACTTA TCCAAAACAA CTTTCCACTG TTGCTTTCGG AGTGAGCAAG ATAGAAAdTG CTCCTTATGT GCAGACAACA TTGAAGGAAA GACATTTGTT TCAACAGTAA ATTCTTTAGT TTTTCAACAA ATAGATGCAA ACTGGAACAT ACAGTGCTGG CTAAAAGGAG ACTTAAAATT ATTCATCTGT TATGTGGAGT CATTATTTAA GAATCTATTC AGGAATTATA ACTATAAGGT CCATCTTTTA TATGTTCTGC CTGAAGTGTT AGAA.GATTCA CCTCTGGTTC CCCAAAAAGG CAGTTTTCAG ATGGTTCACT GCAATTGCAG TGTTCACGAA TGTTGTGAAT GTCTTGTGCC TGTGCCAACA GCCAAACTCA ACGACACTCT CCTTATGTGT TTGAAAATCA CATCTGGTGG AGTAATTTTC CAGTCACCTC TAATGTCAGT TCAGCCCATA AATATGGTGA AGCCTGATCC ACCATTAGGT TTGCATATGG AAATCACAGA TGATGGTAAT CTTGGTCCAG CC CACCATTG GTACCATTTC CACTTCAATA TATTOAGAGA ATTCTACAAC AGTTATCAGA GAAGCTGACA AGCTACATCC CTGCTAGTAG ACAGTATACT TCCTGGGTCT

TTAAAGATTT

TCAAGTGAAA

AGAT TGTCTC

TCGTATGAGG

87- 1201 TTCAGGTCAO GGOCAAGAGA CTGGATGGCO CAGGAATcTG

GAGTGACTGG

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

AGTACTCCTC

AATTOTGACA

AGGAAAACAA

GCT GAGAAAA

CAAAGTTACT

COTATGATC

GOTGAATTAT

TGGGTACTTA

CACTTGGA

TGTTCTGATA

TTTGCAGAGT

TATOTGGCTA4

GACTOTCCAO

TCCATCCAGT

TATOTTGGGA2

CGCTATGGTTI

TGATGCAAA

I

TCTATGCTGT

TI

AGTAATTGGA

G

TATGAGAGGA

C

AGGAGAAAAA

T

TTGTGCAGTG

T

AACATGGTCAG

CAGAGCAAGC

A

TCTGTTGCAA

A

TATCGTGCAG

T(

OTOTOT TTAO

AGTGTTGGGT

CATTOT TCO

TTCCTCAAAG

TTTTTCAATC

AGTGTACTGC

ATGTGATTGA

ACTAAAATGA

AAGCAOTTTG

TTCCATCTAT

GATGGTTTTT

CACAATGTGG

CAACATGTGT

3TGAAAGCAG

%AAGCCAGTC

AAGTGGAAA2 'CAAAATCTG

I

CAGGTGCGC I1 ;CAATCCAGC

C

CTGAATTTT

G

GTCACTTTA C TOAGAGATA

T

AAGATGTGG

G

CATACTGTT

A

TTTTAATTT A.

CACTCAGTG C

CACACAAGAT

CTAATGTTTC

TCAAAAGAGA

CCAGTATGAT

TGAATGAAAO

TGOAATGAAC

TGTCAATATC

O TTGCAGATG

CAATTGAGGT

TCATCCCATA

ATGAATG CAT

ATTAGGATCA

CCTTCCTGAT

kAATTACTAT rTTCCACAGA k~GAAGTACAA

'CAGTCTCCC

~GTAAGAGGC

:TAOACAGTT

C

;GAGAATAAT I1 TTTGGAAGC

C

GTGATAAACC

AAATCACAC

CGGTTCTGG

C

~CCTTTTCA

T

TTATCCTTTA

OTOATATAC

T

TTTTCACTGC

TTGTTTGGTG

GTTGTGAGTG

C.AAACCTCGA

ATGAATGCCA

AATATCTCAT

GTCAACCAGT

ATOATAGGAG

TCTGAGCCCA

TTTCCAGCCA

ATCACTCTCT

TCTGTGGTGA

AAACATTGGA

kTAACCTTCA rGGAAGATGT

"GTTCCAGAC

LAGATGGACT

TCATGGATA

AATGGAGAT

:CCTGATGAA

:ATCATACTT

C

'AAATTCACT

TI

CATCAATTC

GGCCTATGA

G

AACAGCAGT

T

TTCCACCTAA

ATCTATAAGA

CATGAATTTA

ATCATGTTAG

GGAAAGTTTA

TCATOGOTAT

GTGAAACTGA

ACAATCCAGT

CAGCCTTTAC

AAGATTGCTA

ATCTTCCTAT

AGGTTCACTT

AGCCACTGCC

TTAT TGAAAA

PTTCCAGATT

kTGAGGTTTA

ETGTGTGCAG

3GGATATTGG EAAAAGT

TOO

~CTATGAAAA

LAATGACTCA

:CTGCAATGG

'TOOTGTGGA

ATTGGTGCT

CAAAGTAAA

GTGTGATTG

88 2551 2601 2651 2701 .0 2751 2801 2851 2901

TTTCCTGGAT

GAGTGGAAAA

TTCATCTGTT

AGTACCAGTT

AAGATAATTA

TGCAGGTGAC

AGATACATTG

ATGC TTTATT

ACTATCACCO

ATCTTA-ATGA

AAGAAG TATT

CAGTCTTTAC

ATACTT TCAC

TACAAGGACG

ATGAGT TTGG TG TGAAATT T

AGTGATTACA

AGATGGTGAA

ATATCCATGA

CCAATATTTA

TCAAGATGAT

ACGATGACAA

ACAACCCACA

GTGATGCTAT

AGO TAATGTA

ATAAAATGGC

TCATTTTATC

TGGAAGGAGT

ATTGAAAAAC

GTAGGGATCO

ACTAGAATGC

TGCTTTATTT

TTTTATTATT

TTAGAATCTC

CCCATTGAGA

GGGAAAACCA

ACCAGAGTGA

AGACATGATA

AGTGAAAAAA

GTAACCAT

89 Recombinan Han OB Receot-or Protein, Natral So -e Vajn amiro acid secp1Ince (spa, ID. No., 13) 1 MICQKFCVVI 51 101 151 2021 251 301 351 401 451 501 551 601 651.

701 751 801

AGLSKNTSN

LCADN IEGKI

LFRNYNYKVII,

P TAKLNDTLL GNLKI SWSSP

GSSYEVQV.G

H-CIYKKENKI

PRGKFTYDAV

TSTIQSLAES

QP IFLLSGyT IGLLK ISWEK PD LCAVYAVQ

GDTMKKEKNV

FTFLWTEQAH

S SCVIVSWIL

FTIL

LHWEFIYVII

NGHYETAVEF

FVSTVNSLVF

ILLYVLPEVLE

MCLKITSGGV

PL VP F L QYQ KRLDGPG IWS VP SKE IVWWM

YCCNEHIECHH

TLQLRYHRSS

MWIRINHSLG

PVFPENNLQF

VRCKRLD GLG

*AFNLSYPITI

*KFNSSGTHF

*QQIDANWNIC

DSP LVP QKG S

IFQSPLMSVQ

VKYSENSTTV

DWS TP RVFT T

NLAEKIPQSQ

RYAELYVI DV LYCSDIPS

IH

SLDSPPTCVL

QIRYGLSGKE

YWSNWSNPAY

WRFKLSCiTE

NLSKTTFHCC

CWLKGDLKLF

FQMVHCNCSV

P I NNKP DP P

IREADKIVSA

QDVIYFPPKI

YDVVS DHVS K

NINISCETDG

P ISEFKDCYL PD SVVKP LP P

VQWKMYEVYD

TVVMD IKVPM

NSTYDYFLLP

FRSEQDpNCS

ICYVESLFKN

HECCE CLV? V LGLHME

ITDD

TSLLVDSILF

LTSVGSNVSF

VTFFNLNETK

YLTKMTCRWS

QSDGFYECIF

SSVKAEITIN

AKSKSVSLPV

RGPEFWRI

IN

TLLWKPLMKN DSLCSVQRYV INHH-TSCNGT

WSEDVGNHTK

TVTVLAINS

I

SF SDYKLMYF GASVANFNLT

FSWPMSKVNI

I IEWKNLNED

GEIKWLRISS

VQ SLSAYP LN

SVKKYYIHGK

90 Human OB Recector Protein, Natural So1ice Varjkanr QNA (Sea. TD No. 14) 21 GCGGOOGCCA GTGTGATGGA TATCTGCAGA A"TOGGOTTT CTCTGCOTTO 51 1021 0 2.51.

201.

252.

301 352.

0 402.

452.

502.

551 601 .0 651 702.

751 801.

851 .0 901 951 1001 1051 1101 *0 1.151 1201

GGTCGAGTTG

TTGTOAAAAA

TAAC TGCGTT

TCTTGOATGC

GOTO TCAAAG

AACCTAAGTT

AOTTTOOACT

TGCAGACAAC

TTTTTCAACA

GACTTAAAAT

CAGGAATTAT

TAGAAGATTC

TGOAATTGCA

AGCCAAAOTC

GAGTAATTTT

AAGCCTGATO

TTTAAAGATT

ATCAAGTGAA

AAGATTGTOT

TTCGTATGAG

GGAGTGACTG

GACCOOG(GA

TTOTGTGTGG

TAACTTGTCA

CACCAAJATTO

AATA T TCAA

TAATTCAAGT

GTTGCTTTCG

ATTGAAGGAA

AATAGATGOA

TATTCATCTG

AAOTATAAGG

AOCT T GGTT

GTGTTCAOGA

AACGACACTC

OCAGCACOT

CACCAT TAGG

TOAAGGTGTA

TTTTGTTAOA

TATCOAATTA

AAOOTATGAC

ATTCGAATGG

GGTACTCACT

GAGTGAGCAA

AGACATTTGT

AACTGGA.ACA

TTATGTGGAG

TCCATO T TTT

CCCCAAAAAG

ATGTTGTGA.A

TCCTTATGTG

CTAATGTCAG

TTTGCATATG

CT TC TOTGAA

TTGGGAATTTI

OTOOTTGGAG

TAO TTCOTTT

ACATTATGAG

TTTOTAAOT T

GATAGAAACT

TTOAAOAGTA

TAOAGTGCTG

TOATTATTTA

ATATGTTOTG

GOAGTTTTOA

TGTOTTGTGO

TTTGAAAATO

TTOAGOOOAT

GAAATOAOAG

GGTAOOATTT

CAGT TATOAG

GAOAGTATAO

AOTGGATGGO

CAOAAAGA

TCTAATGTTT

GTA.AGATGAT

ATTTATGTGA

ATTTAAGTTG

TGOOTGOTGG

AOAGOTGTTG

ATOOAAAAOA

GOTOOTTATG

AATTOTTTAG

GOTAAAAGGA

AGAATOTATT

COTGAAGTGT

GATGGTTOAO

OTGTGOOAAO

ACATCTGGTG

AAATATGGTG

ATGATGGTAA

COACTTCAAT

AGAAGCTGAO

TTOOTGGGTO

CCAGGAATOT

TGTOATATAO

CTTTTCACTG

TCTTGGTOCA GCOACCATT ATATTOAGAG AATTCTACAA CAGOTACATO CCTGCTAGTA GTTOAGGTGA GGGGOAAGAG GAGTAOTOOT CGTGTCTTTA TTTOOAOOTA AAATTCTGAC A.AGTGTTGGG OATOTATAAG AAGGAAAAOA AGATTGTTC GGATGAATTT AGOTGAGAAA ATTOOTOAAA CTCAAAAGAG ATTGTTTGGT GOOAGTATGA TGTTGTGAGT 91. 2.251 GATCATOTTA GCAAAGTTAC TTTTTTCAAT CTGAATGAAA

OOAAACCTCG

1301 AGGAAAGTTT AOOTATGATG

CACTGTACT

1352.

1402.

1451 .0 1501 1551 1601 1651 1.701 1751 1801 1851 1.901 1951 2001 2051 2101 2151 2201 2251 2301 2351 2401 2451 2501

ATCATCCTA

TGTGAAACTG

TAOAATCCAG

GCAGCCTTTA

AAAGATTGCT

AATOTTCCTA

TAGGTTCACT

AAGCOACTGC

ATTATTGAAA

AATTCCAGAT

TATGAGGT

CTTGTGTGCA

TGGGATATTG

ATAAAAGTTC

TACTATGAAA

AAAA.TGACTC

I

TCCTGCAATGC

TTTCOTGTGG

P.

CAATTGGTGC

TI

AGCAAAGTAA

A

TTGTGTGATT

G

~TTTTATTAT

T

CTTAGAATCT

C

TATACTT

TGCTGAATI

ATGGGTACT

TCACTTGCG

CTOT TC TGA

ATTTGCAGA

TTATC TGGC'

TGACTCTCCJ

OTOCATOCAC

ATATCTTGG(

TCGCTATGG1

ATGATGCAA-P

GTCTATGCTC

GAGTAATTGG

TATGAGAGG

%AGGAGAAAA

~TTGTGCAGT

;AACATGGTC

LOAGAGCAAG

'TCTGTTGCA

*TATCGTGCA

TTTCCTGGA

GAGTGGAAA

TTCATCTGT

*A TATGTGATT T A.AOTAAAAT G AAACCACTT T ATTOOATCT.

G TGATGCTTT' T' AOACAATGT( k. CCAACATGTC 3TGTG-AAAGo; 3 AAAAGCCAGI

TTAAGTGGA.P

ATCAAAATCT

TTCAGGTGCG

*AGCAATOCAG

ACCTG.AATTT

ATGTCACTTT

GTTCAGAGAT

AGAAGATGTG

CACATACTGT

AATTTTAATT

GTCACTCAGT

TACTATCAC

AATCTTAATG

TAAGAAG TAT G0 CTC3CAATGA G ATGTOAATA G ACTTGCAGA T GCAATTCAG k TTOATCCCA r TATGAATG, 3 AT TAGGATC 3TCOTTOOTG2

SGAXATTACTI

OTT TOCAGAC

LAAGAAGTACP

GTCAGTCTCC

OTG TAAGAGG 00 TACACAG

T

TGGAGAATAA

ACTTTGGAAG

ATGTGATAAA

GGAAATCACA

TACGGTTCTG

TAACCTTTTC

GCTTATOOTT

CAG TGATTAC A CATGAATGOC .T OAATATCTOA T GGTCAAOOAG o TATOATAGGA T ATOTGACCC k~ TTTTOOAGCO

:AATOAOTOTO

TTOTGTGGTG

~TAAAOATTGG

AATAAOOTTO

ATGGAAGATG

AGTTOOCA

OTAGATGGAC

TGTOATGGAT

TTAATGGAGA

OOOOTGATGA

COATOATACT

OGAAATTOAO

GOOATOAATT

ATGGOOTATG

TAAACAGCAG

AAGATGGTGA

AATAAAATGG

TATATOCATG GTAAGTTTAO 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.

Claims (5)

1. 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 according to (Seq. ID No. 1): 1-896;

22-896 optionally with an N-terminal methionyl residue;

23-896 optionally with an N-terminal methionyl residue;

29-896 optionally with an N-terminal methionyl residue; 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 derivate of any subparts through 2. An OB receptor protein preparation of claim 1 wherein said OB receptor protein having substituted the C-terminal amino acids, beginning at position 799, G K F T I L (Seq. ID No. 13). 3. An OB receptor protein preparation according to claims 1 or 2, wherein the extracellular domain of said OB receptor protein is modified, said modification selected from among: -141 deletion of all or part of the random coil domain; modification of one or both "WSXWS" boxes by substitution 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. 4. A DNA molecule encoding an OB receptor protein according to any of claims 1-3. An expression or cloning vector containing a DNA of claim 4. 6. A prokaryotic or eukaryotic host cell containing the vector of claim 7. A host cell of claim 6 which is an isolated human host cell. 8. A process for producing an OB receptor protein comprised of culturing, under suitable conditions, a host cell according to claim 6, obtaining the OB receptor produced, and optionally preparing a pharmaceutical composition containing said OB receptor. 9. Use of an OB receptor protein according to claims 1-3, or produced by the process of claim 8, for manufacturing a medicament for the treatment of obesity, diabetes, high blood lipid levels, or high cholesterol levels. An OB protein/OB receptor protein complex preparation, containing an OB protein moiety and an OB receptor protein moiety, optionally in a pharmaceutically acceptable formulation, wherein; said OB receptor protein is selected from among those set forth in any of claims 1 to 3; -142- said OB protein moiety is selected from among: a naturally occurring OB protein; and, (ii) a non-naturally occurring OB protein, analog or derivative thereof. 11. 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 protein/OB receptor protein complex preparation of claim 12. A method of claim 11 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. 13. 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 1-3, or produced by the process according to claim 8. 14. Use of an OB protein/OB receptor protein complex preparation, according to claim 10, for manufacturing a medicament for the treatment of obesity, diabetes, high blood lipid levels, or high cholesterol levels. An OB receptor protein preparation according to claim 1 substantially as herein described with reference to example 7. 16. A use according to claim 9 substantially as herein described with reference to example 7. 17. A method according to claim 12 or 13 substantially as herein described with reference to examples 7 or 9. 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. 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 Vai Val Leu Leu His Trp Giu ?he Ile Tyr Phe Leu Glu Asn Arg Ser Ile Glu 145 Leu Gin I Cys C Leu L 2 Pro L 225 Leu G Va Ly Prc Thr Leu Asn Thr Gln 130 Ser Leu ys :ys ,eu :10 eu ly 1 Ile 5 Leu Ala Ala Ser Cys Val 115 Cys Leu Tyr Gly Glu C 195 Met C Met S Leu H Th Se Gi Va. Ly Se Asr Trp Phe Vial 3er :ys :ys er is r Ala r Cys y Leu I Glu Thr Leu Ser Leu Lys Leu 165 Phe Leu I Leu I Val G 2 Met G Ph Met Se Pro 70 Thr Cys Leu Lys Asn 150 Pro 3In Tal ,ys Asr t Prc r Lys 55 Lvs Phe Ala Vai Gly 135 Leu Glu Met Pro Ile i Leu Ser 25 Pro Asn 40 Asn Thr Phe Asn His Cys Asp Asn 105 Phe Gin 120 Asp Leu Phe Arg Val Leu Vai His 185 Val Pro 1 200 Thr Ser G Tyr Pro Ile Thr Pro Trp Arg Ser Ser Ser Cys Ile Gin Lys Asn 3lu L70 :ys 'hr ;ly Let .sp C Thr Asn Ser 75 Phe Tyr Ser Gly Arg Asp Tyr Asn Gly Thr His Ser Glu PhE His Phe Gin Leu Tyr Ser Asp Glu Ile Leu Tyr 155 Asp Asn kla 31 y Tal !35 ;ly Gly Asp Phe 140 Asn Ser Cys Lys Val 220 Lys Asn Lys Ala 125 Ile Tyr Pro Ser Lea- 205 Ile Pro Leu Thr 110 Asn Cys Lys Leu Vai 190 Asn Phe Asp Lys Phe Va Trp Asr Tyr Val Val His 160 Val Pro 175 His Glu Asp Thr Gin Ser Pro Pro 240 Ile Ser 255 L 215 in 30 lu Ile Thr Asn Asp A 245 250 Trp Ser Ser Pro 260 Pro Leu Vai Pro Phe 265 Pro Leu Gin Tyr Gin Vai Lys 270 95 Tyr Ser Giu Asn Ser Thr ?hr Val Ile Arg Giu Ala Asp Ls Ile Val 275 280 285 Ser Ala Thr Ser Leu Leu Val Asp Ser Ile Leu Pro Gly Ser Ser Tyr 290 295 300 Giu Val Gin Val Arg Gly Lys Arg Leu Asp Giy Pro Gly Ile Trp Ser 305 310 315 320 0 Asp Trp Ser Thr Pro Arg Val Phe Thr Thr Gin Asp Vai Ile Tyr Phe 325 330 335 Pro Pro Lys Ile Leu Thr Ser Vai Gly Ser Asn Val Ser Phe His Cys 340 345 350 Ile Tyr Lys Lys Giu Asn Lys Ile Vai Pro Ser Lys Giu Ile Val Trp 355 360 365 0 Trp Met Asn Leu Ala Giu Lys Ile Pro Gin Ser Gin Tyr Asp Val Val 370 375 380 Ser Asp His Vai 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 Glu Cys His His Arg Tyr Ala Giu Leu Tyr Val Ile Asp Vai Asn Ile O420 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 465 470 4 75 480 11 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 15500 505 510 Ile Arg Ile Asn His Ser Leu Gly 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 Val Lys 530 535

540. 96 Ile Gly Leu Ala Giu Ile Thr Ile Asn 545 550 Leu Lys Ile Ser Trp Giu Lys Pro Val Phe Pro Glu Asn As 565 n Se Sei Va Tr; 625 Arg Glu Leu Gly Trp 705 Gly Lys Cys Tyr Trp 785 Phe r GI Ly L GI; 61 Se; Gi' Lys Cys Thr 690 Thr Ala Val Val Phe 770 Leu Ile y Lys s Ser 595 n Val 0 r Asn Pro Asr Ser 675 Trp Glu Ser Asn Ile 755 Ile I Arg I Pro I Gi 58 Va Ar Pr G1 Val 66C Val Ser ln Jal Ile tal le le le u Val GI 0 1 Ser Le g Cys Ly o Ala Ty 63 a Phe Tr 645 Thr Le Gin Ar Glu AsF Ala His 710 Ala Asn 725 Val Gin Ser Trp Glu Trp Ser Ser 790 Glu Lvs .1 S r 0 P u 9 D n Trp Pro Arg 615 Thr Arg Leu Tyr Val 695 Thr Phe Ser I Ile I 7 Lys A 775 Ser V Le Ly Va 60] Let VaI Ile Trp Val 680 Gly Va1 ksn .eu ,eu '60 iSf 'al u Gin s Met 585 1 Pro 1 Asp Val Ile Lys 665 Ile Asn Thr Leu J Ser 745 Ser P Leu A Lys L Ph 57 Ty As Gi1 Met Asr 65C Prc Asn His lal rhr T30 la ro n ys e Gin 0 r Glu p Leu y Leu Asp 635 1 Gly Leu His Thr Leu 715 Phe Tyr E Ser I Glu A 7 Tyr T 795 Leu T I1 Va Cy Gi' 62 Iit AsE Met His Lys 700 Ala Ser ro ~sp Lsp yr yr e Arg 1 Tvr s Ala 605 v Tyr 0 e Lys Thr Lys Thr 685 Phe Ile Trp Leu Tyr I 765 Gly G Ile H Pro I Ty As 59 Va Trl Va Met Asr 67C Ser Thr ksn ?ro ~sn 150 lys ;lu 'is le r Gly 575 p Ala 0 1 Tyr p Ser i Pro Lys 655 Asp Cys Phe Ser Met 735 Ser S Leu I- Ile L Asp H 8 Phe M 815 Leu Lys Ala Asn Met 640 Lys Ser Asn Leu Ile 720 er er et ,ys i s 00 et 805 Tyr Gin Phe Ser 810 Glu Gly Val Lys Pro Lys Ile Ile 825 Asn Ser Phe Thr Gin Asp Asp 830 97 Ile Giu 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 850 855 860 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 Arg Thr Asp Ile Leu 885 890 895 Ser Leu Ile Met Ile Thr Thr Asp Giu Pro Asn Vai Pro Thr Ser Gin 900 905 910 Gin Ser Ile Glu Tyr Lys Ile Phe Thr Phe Arg Arg Giy Ala Asn Leu 915 920 925 Lys Lys Ile Gin Leu Asn Phe Giu Leu Thr Tyr Gly Gly Leu Cys Phe 930 935 940 Arg Thr Asn Arg Cys Val Asn Leu Gly Ser Lys Cys Arg Phe Giu Ser 945 950 955 960 Ser Leu Asp Vai Leu 965 INFORMIATION FOR SEQ ID NO:2: SEQUENCE CHARACTERISTICS: LENGTH: 3193 base pairs TYPE: nucleic acid STRANDEDNESS: single TOPOLOGY: linear (ii) MOLECULE TYPE: cDNA (xi) SEQUENCE DESCRIPTION: SEQ ID NO:2: CCGCCGCCAT CTCTGCCTTC GGTCGAGTTG GACCCCCGGA GTAAGATGAT TTGTCAAAAA TTCTGTGTGG TTTTGTTACA TAACTGCGTT TA-ACTTGTCA TATCCAATTA CTCCTTGGAG CACCAAATTC AACCTATGAC TACTTCCTTT TGCCTGCTGG ATTCGA-ATGG ACATTATGAG ACAGCTGTTG AACCTAAGTT. TCAAGGTGTA CTTCTCTGAA TTGGGAATTT ATTTATGTGA ATTTAAGTTG TCTTGCATGC ACTCTCAAAG AATACTTCAA TAATTCAAGT GGTACTCACT 120 180 240 300 98 TTTCTAACTT ATCCAAAACA GCTCCTTATG TGCAGACAAC TTTTTCAACA AATAGATGCA TATTCATCTG TTATGTGGAG TCCATCTTTT ATATGTTCTG 0 GCAGTTTTCA GATGGTTCAC CTGTGCCAAC AGCCAAACTC GAGTAATTTT CCAGTCACCT CACCATTAGG TTTGCATATG GCCCACCATT GGTACCATTT 0 CAGTTATCAG AGA.AGCTGAC TTCCTGGGTC TTCGTATGAG GGAGTGACTG GAGTACTCCT AAATTCTGAC AAGTGTTGGG AGATTGTTCC CTCAAAAGAG 0 GCCAGTATGA TGTTGTGAGT CCAAACCTCG AGGAAAGTTT ATCATCGCTA TGCTGAATTA I ATGGGTACTT AACTAAAATG AAAGCACTTT GCAATTGAGG TI 0 TTCATCCCAT ATCTGAGCCC TTTTCCAGCC AATCTTCCTA TI TAGGTTCACT TGACTCTCCA C CTCCATCCAG TGTGAAAGCA G AAAAGCCAGT CTTTCCAGAG A 0 AAGAAGTACA ATGGAAGATG T ACTTTCCACT ATTGAAGGAA GTTGCTTTCG AGACATTTGT GAGLGA-C.A TTCA.ACAGTA GATAGAAACT AATTCTTTAG AACTGGAACA TACAGTGCTG GCTAAAAGGA GACTTAAAAT TCATTATTTA CCTGAAGTGT TGCAATTGCA AACGACACTC CTAATGTCAG GAAATCACAG CCACTTCAAT AAGATTGTCT GTTCAGGTGA CGTGTCTTTA rCTAATGTTT kTTGTTTGGT 3ATCATGTTA kCCTATGATG ~ATGTGATTG LCTTGCAGAT 'ATCATAGGA C LAAGATTGCT 'TATCTGGCT A~ CAACATGTG T AAATTACTA TI ATAACCTTCA ATGAGGTTT A AGAATCTATT TAGAAGATTC GTGTTCATGA TCCTTATGTG TTCAGCCCAT ATGATGGTAA ATCAAGTGAA CAGCTACATC GGGGCAAGAG CCACACAAGA CTTTTCACTG GGATGALATTT GCAAAGTTAC CAGTGTACTG kTGTCAATAT 3GTCAACCAG ;CAGccTTTA C LTTTGCAGAG TI LCACAATGTG G CCTTCCTGA T AAACATTGG A ATTCCAGAT T TGATGCAA.A A CAGGAATTAT ACCTCTGGTT ATGTTGTGAA TTTGAAAATC AA.ATATGGTG TTTAA.AGATT ATATTCAGAG CCTGCTAGTA ACTGGATGGC TGTCATATAC CATCTATAAG PGCTGAGAAA TTTTTTCAAT 2TGCAATGAA 'AATATCTCA L'ACAATCCAG :TGTTCTGAT ~GATGGTTTTI ;ATTAGGATC 'TCTGTGGTG .TTATTGAAA A, CGCTATGGT TI TCAAAATCT G A-AC'TATAAGG CCCCAAAAAG TGTCTTGTGC ACATCTGGTG AAGCCTGATC TCTTGGTCCA AATTCTACAA GACAGTATAC CCAGGAATCT TTTCCACCTA AAGGAAAACA ATTCCTCAAA CTGAATGAA.A CATGAATGCC rGTGAAACTG I'CACTTGCGG k.TTCCATCTA LATGAATGCA L.ATCACTCTC LAGCCACTGC LTATCTTGGG TAAGTGGAA TCAGTCTCC 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 CAGTTCCAGA CTTGTGTGCA GTCTATGCTG TTCAGGTGCG CTGTAAGAGG CTAGATGGAC 1920 99 TGGGATATTG CTATGAGAGG ATGTCACTTT ATGTGATAA.A GAGTA.ATTGG ACCTGAATTT ACTTTGGAAG CCATCATACT .0 CGAAATTCAC TTTCCTGTGG CAATTGGTGC TTCTGTTGCA ATATCGTGCA GTCACTCAGT TACTATCACC CAGTGATTAC AAGATGGTGA AATAAAATGG ?0 ATCATTTTAT CCCCATTGAG TGGGAAAACC A.AAGATAATT ATGCAGGTTT ATATGTAATT CATTATTAAT ATCACACCAA AGAATTGTTC CTGGGCACAA TAATCATGAT CACTACAGAT AGAAGATTTT TACATTTTGA GAGAGTTAAC ATATGGTGGA GTTCAA.AATG TAGATTTGAG TTTAAAAGTA GTATTCATGA TTAAGAAAAT TATGGCTGTT AGCAATCCAG TGGAGAATAA CCCCTGATGA TCCTGCAATG ACAGAGCAAG AATTTTAATT GCTTATCCTT AAGCTAATGT CTTAGAATCT AAGTACCAGT AATAGTTTCA STGCCAGTAA 3GACTTAATT 3AACCCAATG k.GAAGGGGAG 7'TATGTTGAT 'CCAGTTTGG 'TTCTGGCTT IJ ~CTGTCATTA C CCTACACAGT TTAATGGAGA AAAATGACTC GAACATGGTC CACATACTGT TAACCTTTTC TAAACAGCAG ATTTTATTAT CTTCATCTGT TCAGTCTTTA CTCAAGATGA TTATTTCCTC AGCTATTTTG TTCAGAAGAG TGCCAACTTC CAAATCTAAA rTAGAACTTA k.TGTGTGATT ~TGATTTGCC I ATATCTATT TGTCATGGAT TACTATGAAA ATTGTGCAGT AGAAGATGTG TACGGTTCTG ATGGCCTATG TTGTGTGATT TGAGTGGAAA TAAGAAGTAT CCCAATATTT TAT TGAA.AAA TTCCATCTTA GGA.AGATGTT AACGGACATT :CAACAGTCT kAAAATTCAG k.AATAGATGT ~aTTTTCAAA LTATTCCTGG LAATGTCATC ATAAAAGTTC AAGGAGAAAA GTTCAGAGAT GGAA.ATCACA GCCATCAATT AGCAAAGTA.A GTTTCCTGGA AATCTTAATG TATATCCATG ATGGAAGGAG CACCAGAGTG TTGCTTGGAA CCGAACCCCA CTTTGAAGTC ATAGAGTATT TTGAACTTCT GTAAATTTGG rCATCTAAAG LCATAAAACA LXATATGTAG 1980 2040 2100 2160 2220 2280 2340 2400 2460 2520 2580 2640 2700 2760 2820 2880 2940 3000 3060 3120 3180 3193 TAGACAATTT TGTAATTAGG TGAACTCTAA AAcTGCAACA TCTGACAAAT TGCTTTAAAA 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 Val Leu Leu His Trp GJlu Phe Ile I1 Tyr Phe Leu Val1 Lys Pro Ii' Le Ala Giu Thr Ala Asn Arg Ser Ile Glu 145 Leu Gin Cys Leu Leu Asn Th r Gin 130 Ser Leu Lys ys Leu Ser Cys Val 115 Cys Le u Tyr Gly Giu 195 Met *Thr *i Ser G.1y IVal Lys Ser 100 As n Trp Phe Val Ser E 180 Cys I Cys L Al C y Let Git Thr Leu Ser Leu Lys Leu 165 ~he ~e u a Phe s Met i Ser 1Pro 70 Thr Cys Le u Lys Asn 150 Pro Gin b Val E Lys I Asr P rc Lys 55 Lys P he Al a Val1 Gly 135 Leu Glu e t Lei Pr 40 Asr Phe His Asp Phe 120 Asp Phe Val1 Val Ser Tyr Pro 25 Asn Ser Thr Thr Ser Asn Asn Ser Ser 75 Cys Cys Phe 90 Asi Ile Giu 105 Gin Gin Ile Leu Lys Leu Arg Asn Tyr 155 Leu Giu Asp 170 His Cys Asn 185 Pro Thr Ala I Ser Gly Gly Ty Se GIj Arc Gly Asp Phe 140 As n Ser .'ys Asp "Asn Thr Ser Lys Aila 125 Ile Tyr Pro Ser Leu 205 Ty r Gly His Giu Th r 110 Asn Cys Lys Leu lai ~sn2 P he His Phe Gin Phe T rp Tyr Val1 Ile Thr Pro Trp Arg Le u Tyr Ser Asp Vali As n Val1 His Val1 175 .iis ksp Pro Giu Thr Ser Val 200 Thr 1210 215 ra 1 lie P'he C;ln 220 Pro 225 Leu Met Ser Val Gin 230 Pro Ile Asn Met Val Lys 235 Pro Asp Pro Pro 240 LO L T SE GI As Pr 11 Trj Se: 38~ Prc Glu Asn Trp Arg 465 Pro Glu Ile eu Gly rp Ser ?r Ser ~r Ala 290 .u Val '5 p Trp o Pro e Tyr p Met 1 370 r Asp H Arg G Cys H Ile S 4 Ser Ti 450 Tyr H Ile Se Cys Ii Arg Ii 51 L SE GI 27 Th Gi Se Ly. Ly 35~ sr Li nly is er 35 ir e 5 eu His Met 245 ar Pro Pro 260 .u Asn Ser p 5 r Ser Leu n Val Arg r Thr Pro 325 s Ile Leu 340 s Lys Glu i Leu Ala C Val Ser I 3 Lys Phe T 405 His Arg T 420 Cys Giu T Ser Thr I Arg Ser SE 4- Glu Pro L 3 485 Phe Gin Pr 500 Asn His Se Glu I Leu V Thr T Leu V 25 Gly Lj 310 Arg Va Thr Se Asn Ly 3lu Ly. 37. ~ys Va 190 hr Tyi yr Ala hr Asp le Gin 455 ar Leu 70 rs Asp :o Ile r Leu 101 le Thr Asp al Pro Phe 265 hr Val Ile 280 ai Asp Ser 95 's Arg Leu .i Phe Thr r Val Gly 345 s Ile Val E 360 s Ile Pro G 5 1 Thr Phe Asp Ala V 4 Glu Leu T' 425 Gly Tyr L 440 Ser Leu A: Tyr Cys SE Cys Tyr Le 49 Phe Leu Le 505 Gly Ser Le 520 A: P2 PI Ar Ii As Th 33 Sec ;ir he al 10 yr La rr uu 0 u u sp Gly 50 :o Leu :g Glu e Leu p Gly 315 r Gin 0 r Asn Ser I Ser C Asn L 395 Tyr C Vai I Thr L Glu SE 41 Asp IJ 475 Gin Se Ser Gi Asp Se A G A P1 3C P I As V a 18' ie y 3 e r Le r y r sn Leu in Tyr la Asp 285 ro Gly o Gly p Vai 1 Ser s Glu 365 r Tyr P u Asn G 5 Cys A Asp V 4 Met TI 445 Thr L Pro SE Asp Gl Tyr Th 51 Pro Pr 525 LI GI 27 Ly Se Ii III ?he 35 Cle lu sn al hr BU rr rr 0 0 ys I 2. .n V~ s5 I r Se e Tr e Ty 33 Hi 0 Va. Va Thi Glu 415 Asn Cys Gin Ile Phe 495 Met Thr le Ser al Lys -e Val r Tyr p Ser 320 r Phe s Cys 1 Trp L Val Lys 400 His Ile Arg Leu His 480 Tyr Trp Cys 102 Vai Leu Pro Asp Ser Val Val Lys Pro Leu Pro Pro Ser Ser Val. Lys 530 535 540 Ala Giu Ile Thr Ile Asn Ile Giy Leu Leu Lys Ile Ser Trp Giu Lys 545 550 555 560 Pro Val Phe Pro Giu Asn Asn Leu Gin Phe Gin Ile Arg Tyr Giy Leu 565 570 575 0 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 Ala 595 600 605 Vai Gin Val Arg Cys Lys Arg Leu Asp Giy Leu Gly Tyr Trp Ser Asn 610 615 620 0 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 Giu 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 Vai Ile Asn His His Thr Ser Cys Asn 0 675 680 685 Gly Thr Trp Ser Giu Asp Val Gly Asn His Thr Lys Phe Thr Phe Leu 690 695 700 Trp Thr Glu Gin Ala His Thr Val 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 0 Lys Val Asn Ile Val Gin Ser Leu Ser Ala Tyr Pro Leu Asn Ser Ser 740 745 750 Cys Val Ile Val Ser Trp Ile Leu Ser Pro Ser Asp Tyr Lys Leu Met 755 760 765 Tyr Phe Ile Ile Glu Trp Lys Asn Leu Asn Glu Asp Gly Glu Ile Lys 770 775 780 0 Trp Leu Arg Ile Ser Ser Ser Val Lys Lys Tyr Tyr Ile His Asp His 785 790 795 800 P he Giu Ile Giy Pro Val1 Ile Gly Giu 805 Lys 103 Tyr Gin Lys Ile P he Ser 810 As n La u Se: Tyr P he Pro Thr Ile Phe 815 Met 820 825 830 Ile Giu Lys His Gin Ser Asp Ala Giy 835 840 Ilec Gin 865 Cys Phe Leu Lys Leu 945 Se: Sle 850 *Arg Se r La u Lys Met 930 Lys Leu Ser Met T rp Ser Gin 915 Arg PArg Arg Ser Lys Ala Se: 900 Phe Cys Val1 Leu Ser Lys Gin 885 Ile Gin Gin Leu Arg Ile Leu 870 Giy Gin Lys Gin P he 950 V1al Le u 855 Phe Leu His Ile Le u 935 Val1 Leu T rp As n Gin Ser 920 T rp Leu La u Giu Phe His 905 ValI Ser Val Leu Tyr Giy Thr Asp Val 875 Gin Lys 890 Val Vai Leu Ile Leu Tyr Thr Ser Val1 Leu 860 Pro Lys Le u His Phe 940 845 Leu As n Arg P he His 925 Gin *Vai Ile Pro La u Phe 910 Gly Gin P rc Ser Lys Se r 895 T rp Lys Glm Val His As n 880 Ile Ser Ile Ile Se: Thr Val ~eu Thr 955 960 Leu Arg Pro Met Arg Thr Lys Aia Arg Asp 970 975 965 Asn Pro Leu Leu Asn Thr Pro Arg 980 Lau Vai Lys 995 INFORMATION FOR SEQ ID NO:4: SEQUENCE CHARACTERISTICS: LENGTH: 3063 base pairs TYPE: nuclij acid STRANDEDNESS: single TOPOLOGY: linear (ii) MOLECULE TYPE: cDNA Ser 985 Ala Thr Leu Asn Gin Val Lys 990 104 (xi) SEQUENCE DESCR.IPTION: SEQ ID NO:4: CCGCCGCCAT CTCTGCCTTC GGTCGAGTTG GACCCCCGGA TCAAGGTGTA CTTCTCTGA.A GTA.AGATGAT TTGTCAAA TAACTGCGTT TAACTTGTC CACCAAATTC AACCTATGA ATTCGAATGG ACATTATGA TTTCTAACTT ATCCAAAAC. GCTCCTTATG TGCAGACAA( TTTTTCAACA A.ATAGATGC TATTCATCTG TTATGTGGAC 0 TCCATCTTTT ATATGTTCTC GCAGTTTTCA GATGGTTCAC CTGTGCCAAC AGCCAAACTC GAGTAATTTT CCAGTCACC7 CACCATTAGG TTTGCATATG 0 GCCCACCATT GGTACCATTT CAGTTATCAG AGAAGCTGAC TTCCTGGGTC TTCGTATGAG GGAGTGACTG GAGTACTCCT AAATTCTGAC AAGTGTTGGG 0 AGATTGTTCC CTCAAAAGAG GCCAGTATGA TGTTGTGAGT CCAAACCTCG AGGAAAGTTT ATCATCGCTA TGCTGAATTA TTCTGTGTGG TTTTGTTACA TTGGGAATTT ATTTATGTGA A TATCCAATTA C TACTTCCTTT G ACAGCTGTTG A~ ACTTTCCACT ZATTGAAGGAA AACTGGAACA 3TCATTATTTA ;CCTGAAGTGT TGCAATTGCA AACGACACTC CTAATGTCAG GAAATCACAG CCACTTCAAT AAGATTGTCT C GTTCAGGTGA C CGTGTCTTTA C TCTAATGTTT C ATTGTTTGGTG GATCATGTTA G ACCTATGATG C TATGTGATTG A CTCCTTGGAG TGCCTGCTGG AACCTAAGTT GTTGCTTTCG AGACATTTGT TACAGTGCTG AGAATCTATT TAGA.AGATTC GTGTTCATGA TCCTTATGTG rTCAGCCCAT ~.TGATGGTA k.TCAAGTGAA AGCTACATCC ;GGGCAAGAG I~ :CACACAAGA T TTTTCACTG C GATGAATTTA CAAAGTTAC T AGTGTACTG C TGTCAATAT C. ATTTAAGTTG ACTCTCAAAG TAATTCAAGT GAGTGAGCAA TTCAACAGTA GCTAAAAGGA CAGGA.ATTAT ACCTCTGGTT ATGTTGTGAA TTTGAAAATC ~AATATGGTG rTTAAAGATT kTATTCAGAG ~CTGCTAGTA ~CTGGATGGC GTCATATAC ATCTATAAG .GCTGAGAAA TTTTTCAAT C TGCAATGAAC AATATCTCA T PCAATCCAG T TCTTGCATGC AATACTTCA.A GGTAC'TCACT GATAGAAACT A.ATTCTTTAG GACTTAAAAT A-AC TAT AAGG CCC CA.AAAAG TGTCTTGTGC ACATCTGGTG AAGCCTGATC TCTTGGTCCA PATTCTACAA GACAGTATAC :CAGGAATCT L'TTCCACCTA L.AGGAAAACA LTTCCTCAAA :TGAATGAAA ~ATGAATGCC GTGAAACTG CACTTGCGG 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 ATGGGTACTT AACTAAAATG ACTTGCAGAT GGTCAACCAG AAAGCACTTT TTCATCCCAT GCAATTGAGG ATCTGAGCCC TATCATAGGA AAAGATTGCT GCAGCCTTTA ATTTGCAGAG CTGTTCTGAT ATTCCATCTA TGATGGTTTT TATGAATGCA 105 TTTTCCAGCC AATC'TTCCTA TAGGTTCACT TGACTCTCCA CTCCATCCAG TGTGAA.AGCA AAAAGCCAGT CTTTCCAGAG 0 AAGAAGTACA ATGGAAGATG CAGTTCCAGA CTTGTGTGCA TGGGATATTG GAGTAATTGG CTATGAGAGG ACCTGAATTT ATGTCACTTT ACTTTGGA.AG 0 ATGTGATAAA CCATCATACT CGAALATTCAC TTTCCTGTGG CAATTGGTGC TTCTGTTGCA ATATCGTGCA GTCACTCAGT TACTATCACC CAGTGATTAC ~0 AAGATGGTGA A.ATAAAATGG ATCATTTTAT CCCCATTGAG TGGGAAAACC AAAGATAATT ATGCAGGTTT ATATGTAATT CATTATTAAT ATCACACCAA AGAATTGTTC CTGGGCACA.A CAAGCATACA GCATCAGTGA AGATATCAGT GTTGATACAT CTCTCTACTT TCAACAACAG CAGTGTTAAC TTCTCTGAGG ACAACCCTTT GTTAAATACG AGA TTATCTGGCT CCA.ACATGTG GAAATTACTA A.ATAACCTTC TATGAGGTTT GTCTATGCTG AGCAATCCAG TGGAGAATA.A CCCCTGATGA TCCTGCAATG ACAGAGCAAG AATTTTAATT GCTTATCCTT AAGCTAATGT CTTAGAATCT AAGTACCAGT AATAGTTTCA GTGCCAGTAA AGAATGAAAA GGACTTAATT CATGTGGTCC CATGGAAAA ATCTTGAAAA CTGAGGGTAC CCACGCTGAT ACACAATGTG TCCTTCCTGA TAAACATTGG A.ATTCCAGAT ATGATGCAAA TTCAGGTGCG CCTACACAGT TTA.ATGGAGA AAAATGACTC GAACATGGTC CACATACTGT TA.ACCTTTTC TAAACAGCAG ATTTTATTAT CTTCATCTGT TCAGTCTTTA CTCAAGATGA TTATTTCCTC AGCTATTTTG TTCAGAAGAA TCTTCTTTTG TAA.AGATGAG GGGTTCTGTT TGAGGTAACC CAGCAACTCT GAT TAGGATC TTCTGTGGTG ATTATTGAAA TCGCTATGGT ATCAAA.ATCT CTGTAAGAGG TGTCATGGAT TACTATGAAA ATTGTGCAGT AGA.AGATGTG TACGGTTCTG ATGGCCTATG TTGTGTGATT TGAGTGGAAA TAAGAAGTAT CCCAATATTT TATTGAAAAA TTCCATCTTA GGAAGATGTT ACGTTTGAGC GAGCCTGAAA ATGATGCCAA TGTTTTAGTG TATGAGGACG A.AACCAAGTG XATCACTCTC AAGCCACTGC ATATCTTGGG TTALAGTGGAA GTCAGTCTCC CTAGATGGAC ATAAAAGTTC AAGGAGAAAA GTTCAGAGAT GGAAATCACA GCCATCAATT AGCAAAGTAA GTTTCCTGGA AATCTTAATG TATATCCATG ATGGAAGGAG CACCAGAGTG TTGCTTGGAA CCGAACCCCA ATCTTTTTAT CAATTTCAGA CAACTGTGGT ACCAGTTCAA AAAGCCAGAG AA.AC'TGGTGA 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 Ci) SEQUENCE CHARACTERISTICS: LENGTH: 969 amino acids TYPE: amino acid STRANDEDNESS: single TOPOLOGY: linear (ii) MOLECULE TYPE: protein (xi) SEQUENCE DESCRIPTION: SEQ ID M'et Ile Cys Gin Lys Phe Cys Val Vai Leu Leu His Trp Giu Phe Ile 1 Tyr P he Leu Giu Asn Arg Ser Ile Giu 145 Leu Gin Val2 Lys P rc Th r Le u As n Th r Gin 130 Ser Leu Lys LIle Thr Leu Ser Ala Gly Ala Val Ser Lys Cys Ser 100 Val Asn 115 Cys Trp Leu Phe Tyr Val Gly Ser 180 Al a Cys Leu Giu Thr Leu Ser Leu Lys Lieu 1.65 'he ?he Met Se r Pro 70 Th r Cys Leu Lys Asn 150 Pro Gin b As r Prc Lys 55 Lys Phe Ala Val1 Gly 135 Leu 31u et Leu Pro 40 As n Phe His Asp P he 120 Asp Phe Val Val I Ser 25 As n Th r As n Cys As n 105 Gin Leu .Arg .eu 'is Tyr Ser Ser Se r Cys 90 Ile Gin Lys Asn Glu 170 Cys Pro Thr As n Ser 75 P he Giu Ile Leu Ty r 155 Asp As n le Tyr Ser Gly Arg Gly Asp Phe 140 As n Ser Cys Th r Asp As n Thr Ser Lys Ala 125 Ile Tyr Pro Ser Pro Tyr Gly His Giu Thr 110 Asn. Cys Lys Leu Val 190 Trp P he His Phe Gin Phe T rp Tyr Val1 Val1 175 Hiis Arg Leu Tyr Ser Asp Val1 As n Val1 His 160 Pro Giu 185 Cys Cys Giu 195 Cys Leu Vai Pro Val 200 Pro Thr Ala Lys Leu Asn Asp Thr 205 Leu Pro 225 Leu Trp Tyr Ser Glu 305 Asp Pro E Ile T Trp M 3 Ser A 385 Pro A Glu C' Asn. I Trp SE Arg Ty 465 LE 21 Le Gi Se Se Al 29 Va rrp ro yr let 70 sp rg ys Le r0 rr ~u Mt .0 tu ME y Le r Se r GI 27 a Th 0 L Gli Sei Lys Lys 355 Asn His Gly His Ser 435 Thr His et Cys 't Ser *u His :r Pro 260 u As 5 r Ser Val Thr E Ile L 340 Lys G Leu A Val S Lys P1 4( His A~ 420 Cys GI Ser Th Arg Se L 2 P1 Se Le rr ~2. la er he 05 :g uu rr r eu Lys al Gin 230 at Glu :o Leu r Thr u Leu g Gly 310 o Arg 5 u Thr Asn I Glu L 3 Lys V 390 Thr T Tyr A. Thr A Ile G2 Ser Le 470 I: 2: P2 I Va Th Va 29. Ly: Val Ser ~ys lys 175 al yr la sp Ln 5S uu 107 Le Thr Se L5 *0 Ile As .e Thr As i Pro Ph 26] r Val IlE 280 1 Asp Ser 5 s Arg Leu Phe Thr Vai Gly 345 Ile Vai 360 Ile Pro Thr Phe Asp Ala Glu Leu 425 Gly Tyr 440 Ser Leu Tyr Cys rG n. Me p A., e P Ar Il As Th 33' Se Prc Gir Phe Vai 410 Tyr Leu Ala 'er ly Gly et Val 235 3p Gly 50 :o Leu g Glu e Leu p Gly I 315 r Gin I 0 r Asn V Ser L Ser G 3 Asn L 395 Tyr C Val I2 Thr L Glu Se 46 Asp I 475 Ve 2; Ll As Gi Al Pr Pr ~SF 'al ys In eu FS Le 0 e l lie s5 Pro ;n Leu n Tyr a Asp 285 o Gly o Gly Vai Ser P 3 Glu I 365 Tyr A Asn G Cys A: Asp V 4: Met Th 445 Thr Le Pro Se Ly Gl 27 Ly Se Li :le he le sp lu sn Li r ie G 5p PJ 's I n Va 0 5 Ii r Se a Tr Ty 33] Hi. Val Va1 Thr Glu 415 Asn Cys Gin Ile in Ser ro Pro 240 .e Ser i Lys e Val r Tyr p Ser 320 r Phe 3 Cys Trp Vai Lys 400 His Ile Arg Leu His 480 108 Pro Ile Ser Glu Pro Lys Asp 485 Glu Ile Val2 Al a 545 Pro Ser Se r ValI Trp 625 Arg Giu Leu Gly T rp 705 Gly Lys Cys Cys Arg Leu 530 Giu Val1 Gly Lys Gin 610 Ser Gly Lys Cys Th-r 690 Thr Ala Val Val1 Ile Ile 515 Pro Ile Phe Lys Se r 595 Val1 As n Pro As n Ser 675 Trp Giu Ser Asn Ile 755 Phe 500 As n Asp Th r Pro Glu 580 Val1 Arg Pro Giu Val1 660 Val1 Ser Gin Val Ile 740 Val1 Gin His Se r Ile Giu 565 ValI Ser Cys Al a P he 645 Thr Gin Giu Ala Al a 725 Val. Ser Pro Ser Val2 As n 550 As n Gin Le u Lys Tyr 630 Trp Leu Arg Asp His 710 Asn Gin T rp Ile Leu Val1 535 Ile As n T rp Pro Arg 615 Th r Arg Leu Ty r Val1 695 Thr Phe Ser Ile Cys P he Giy 520 Lys Gly Leu Lys Val1 600 Leu Val Ile Trp Vali 680 Giy Val Asn Leu Leu 760 490 495 Tyr Leu Gin Ser Aso Giy ?he Tyr Le u 505 Se r Pro Leu Gin Met 585 Pro Asp Val1 Ile Lys 665 Ile Asn Thr Leu Ser 745 Ser Le u Leu Leu Leu Phe 570 Tyr Asp Gly Met As n 650 Pro Asn His Val Thr 730 Ala Pro Ser Asp Pro Ly s 555 Gin Giu Leu Leu Asp 635 Gly Leu His Tkir Leu 715 P he Tyr 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 Aia 605 Tyr Lys Th r Lys 'Thr 685 Phe Ile Trp Leu Tyr 765 Thr 510 Pro Ser T rp Tyr Asp 590 Val2 T rp Val Met Asn 670 Ser Thr Asfl Pro Asn 750 Lys Met Th r Val2 Glu Giy 575 Ala Tyr Ser Pro Lys 655 Asp Cys Phe Ser Met 735 Se r Leu T rp Cys Ly s Lys 560 Leu Ly s Ala As n Met 640 Lys Ser Asn Leu Ile 720 Ser Ser Met 109 Tyr Phe Ile Ile Giu Trp Lys Asn Leu Asn Giu Asp Gly Giu Ile Lys 770 775 780 Leu Arg Ile Ser Ser Ser Val 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 ?he Met 805 810 815 Glu Gly Val Giy 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 Val Ile Val Pro Val 5835 840 845 Ile Ile Ser Ser Ser Ile Leu Leu Leu Giy Thr Leu Leu Ile Ser His 850 855 860 0Gin Arg Met Lys Lys Leu Phe Trp Giu Asp Val Pro Asn Pro Lys Asn 865 870 875 880 Cys Ser Trp Aia 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 Gly 900 905 910 His Lys His Cys Giy Arg Pro Gin Giy Pro Leu His Arg Lys Thr Arg 0915 920 925 Asp Leu Cys Ser Leu Vai Tyr Leu Leu Thr Leu Pro Pro Leu Leu Ser 930 935 940 Asp Pro Ala Lys Ser Pro Ser Vai Arg Asn Thr Gin Giu Ser Ile 945 950 955 960 Lys Lys Lys Lys Lys Lys Leu Giu Giy 965 INFORMATION FOR SEQ ID NO:6: SEQUENCE CHARACTERISTICS: LENGTH: 969 amino acids TYPE: amino acid STRANDEDNESS: single TOPOLOGY: linear (ii) MOLECULE TYPE: protein 110 (xi) SEQUENCE DESCRIPTION: SEQ ID NO:6: Met Ile Cys Gin Lys Phe Cys Val Val Leu Leu His Trp Giu Phe Ile Ty PhE Let GIL Asn Arg Ser Ile Glu 145 Leu Gin Cys Leu Pro 225 Leu r Vai B Lys i Pro Thr Leu Asn Thr Gin 130 Ser Leu Lys Cys Leu b 210 Leu 1 Gly I I I Let Al Al Ser Cys Val 115 Cys Leu Tyr ly lu 195 4et let leu Th~ Sei a G13 i Val Lys Ser 100 Asn Trp Phe Vai Ser 180 Cys Cys Ser His r Ala Cys r Leu Glu Thr Leu Ser Leu Lys Leu 165 Phe Leu Leu I Val G 2 Met G Pht Met Ser Pro 70 Thr Cys Leu Lys Asn 150 Pro 3In lal ys ;in 130 ;lu e Asn Pro Lys 55 Lys Phe Ala Vai Gly 135 Leu Glu Met Pro Ile *j 215 Pro I Ile T Let Prc 40 Asr Phe His Asp Phe 120 Asp Phe ial lai lal !00 'hr :le hr u Ser 25 Asn i Thr Asn Cys Asn 105 Gin Leu Arg Leu His 185 Pro Ser C Asn Asp A Ty Sei Sei Sei Cys 90 Ile Gin Lys Asn Glu 170 :Y3 Lhr ;ly let ~sp r Pro Thr Asn Ser 75 Phe Glu lie Leu Tyr 155 Asp Asn Ala Gly Vai I 235 Gly I IlE Tvr Sez Gly Arg Gly Asp Phe 140 Asr Ser :ys Lys lal 220 -ys sn Th Ast Asr Thr Ser Lys Ala 125 Ile Tyr Pro Ser Leu 205 Ile Pro Leu r Pr: Ty GI1 His Glu Thr 110 Asn Cys Lys Leu Vai 190 Asn Phe Asp Lys Tro Phe o His Phe Gin Phe Trp Tyr Vai Val 175 His Asp Gin Pro I Ile S Arg Leu Tyr Ser Asp Val Asn Va1 His 160 Pro lu rhr 3er !ro ;er 250 255 Trp Ser Ser Pro 260 Pro Leu Vai Pro Phe 265 Pro Leu Gin Tyr Gin Vai Lys 270 ill Tyr Ser Giu Asn Ser Thr Thr Val Ile Arg Glu Ala Asp Lys Ile Val 275 280 285 Ser Ala Thr Ser Leu Leu Val Asp Ser Ile Leu Pro Giy Ser Ser Tyr 290 295 300 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 Val 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 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 Vai 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 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 Se r 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 Gly Tyr Thr Met Trp 50.0 505 510 Ile Arg Ile Asn His Ser Leu Gly Ser Leu Asp Ser Pro Pro Thr Cys 515 520 525 Vai Leu Pro Asp Ser Vai Val Lys Pro Leu Pro Pro Ser Ser Val Lys 530 535 540 Ala Giu Ile Thr Ile Asn Ile Gly Leu Leu Lys Ile Ser Trp Giu Lys 545 550 55560 1.12 Pro Val Phe Pro Giu Asn Asfl Leu Gin Phe *i-n Ile Arg Tyr Gly Leu 565 570 575 Ser Gly Lys Giu Val Gin Trp Lys Met Tyr Giu Vai Tyr Asp Ala Lys 580 585 590 Ser Lys Ser Val Ser Leu Pro Val Pro Asp Leu Cys Ala Val Tyr Ala 595 600 605 LO0 Val Gin Val Arg Cys Lys Ar; Leu Asp Gly Leu Gly Tyr Trp Ser Asn 610 615 620 Trp Ser Asn Pro Ala Tyr Thr Vai Vai Met Asp Ile Lys Val Pro Met L 5 625 630 635 640 Arg Giy Pro Giu Phe Trp Arg Ile Ile Asn Gly Asp Thr Met Lys Lys 645 650 655 )0 Giu Lys Asn Val Thr Leu Leu Trp Lys Pro Leu Met Lys Asn Asp Ser 660 665 670 Leu Cys Ser Vai Gin Arg Tyr Vai Ile Asn His His Thr Ser Cys Asn 675 680 685 Gly Thr Trp Ser Giu Asp Vai Gly Asn His Thr Lys ?he Thr Phe Leu 690 695 700 Trp Thr Giu Gin Ala His Thr Vai Thr Vai Leu Ala Ile Asn Ser Ile 705 710 715 720 Gly Ala Ser Vai Ala Asn Phe Asn Leu Thr Phe Ser Trp Pro Met Ser 725 730 735 Lys Vai Asn Ile Val Gin Ser Leu Ser Ala Tyr Pro Leu Asn Ser Ser 740 745 750 Cys Val Ile Vai Ser Trp Ile Leu Ser Pro Ser Asp Tyr Lys Leu Met 755 760 765 Tyr Phe Ile Ile Giu Trp Lys Asn Leu Asn Glu Asp Giy Glu Ile Lys 770 775 780 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 820 825 830 113 lie Glu Lys His Gin Ser Asp Ala Gly Leu Tyr Val Ile Val Pro Val 835 840 845 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 0 Cys Ser Trp Ala Gin Gly Leu Asn Phe Gin Lys Met Leu Glu Gly 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 .0 930 935 940 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 Glu Gly 965 INFORMATION FOR SEQ ID NO:7: 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 Glu Phe Ile 1 5 10 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 35 40 Leu Pro Ala Gly Leu Ser Lys Asn Thr Ser Asn Ser Asn Gly His Tyr 55 114 Glu Asn Arg Ser Ile Glu 145 Leu Gin Cys Leu Pro 225 Leu C Trp S Tyr S Ser A 2 Glu V 305 Th Le As: Th: GIz 13( Se2 Let Lys Cys Leu 210 Au ily ier er la 90 al r Ala u Ser Cys Val 115 Cys Leu Tyr Gly Glu 195 Met Met Leu Ser P Glu P 275 Thr S Gin V Va Ly: Se Asr Trp Phe Val Ser 180 Cys Cys Ser lis 'ro Lsn er al 1 Glu s Thr r Leu i Ser Leu Lys Leu 165 Phe Leu Leu Val Met 245 Pro I Ser I Leu L Arg G Pr Th. Cy~ Let Lys Asn 150 Pro Gin Val Lys Gin 230 3lu .eu :hr leu ly o Lys r Phe s Ala Val Gly 135 Leu Glu Met Pro Ile 215 Pro Ile Val I Thr Val I 295 Phe Asn Ser Ser Gly Thr His Phe Ser Hi, As Ph 12( Asr Phe Va1 Va1 Va1 200 Thr Ile rhr ?ro Tal !80 sp s Cys p Asn 105 Gin Leu Arg Leu His 185 Pro Ser Asn Asp Phe 265 Ile I Ser I Cy 90 Ilt G1i Ly Asr Glu 170 Cys Thr Gly Met Asp 250 ?ro krg :le 5 Phe Arg e Glu Gly n Ile Asp s Leu Phe ,140 I Tyr Asn 155 Asp Ser Asn Cys Ala Lys Gly Val 220 Val Lys 235 Gly Asn I Leu Gin I Glu Ala Leu Pro G 300 Gly Pro G 315 Se: Ly, Ala 125 Ile Tyr Pro Ser Leu 205 Ile Pro Leu yr sp ;ly ily r GI Th, l1( Ast Cys Lys Leu Va1 190 Asn Phe Asp Lys Gin 270 Lys Ser Ile u Gin Phe 0 n Trp 3 Tyr Val Val 175 His Asp Gin Pro Ile 255 Vai Ile Ser I Trp S 3 Asp Val Asn Val His 160 Pro Glu Thr Ser Pro 240 Ser ys fal 'yr er Lys Arg Leu Asp 310 Asp Trp Ser Thr Pro 325 Arg Val Phe Thr Thr 330 Gin Asp Val Ile Tyr Phe 335 115 Pro Pro Lys Ile Leu Thr Ser Val Gly Ser Azn Vai Ser Phe His 340 345 350 Ile Tyr Lys Lys GJlu Asn Lys Ile Val Pro Ser Lys Glu Ile Vai Trp 355 360 363 Trp Met Asn Leu Aia Giu Lys Ile Pro Gin Ser Gin Tyr Asp Vai Val 370 375 380 LO Ser Asp His Vai 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 L Giu Cys His His Arg Tyr Aia Giu Leu Tyr Val 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 Aia 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 485 490 495 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 515 520 1525 Vai Leu Pro Asp Ser Vai Vai Lys Pro Leu Pro Pro Ser Ser Val Lys 530 535 540 Ala Giu Ile Thr Ile Asn Ile Giy Leu Leu Lys Ile Ser Trp Giu Lys 545 550 555 560 Pro Vai Phe Pro Giu Asn Asn Leu Gin Phe Gin Ile Arg Tyr Gly Leu 565 57057 Ser Gly Lys Giu Vai Gin Trp Lys Met Tyr Giu Vai Tyr Asp Aia Lys 580 585 590 Ser Lys Ser Vai Ser Leu Pro Vai Pro Asp Leu Cys Aia Val Tyr Ala 595 600 605 Val Gin Val Ar; Cys Lys Arg Leu Asp Giy Leu Gly Tyr Trp Ser Asn 610 615 620 116 Trp Ser Asn Pro Ala Tyr Thr Val Val Met Asp Ile Lys Val Pro Met 625 630 635 640 Arg Gly Pro Glu 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 0 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 Val Gly Asn His Thr Lys Phe Thr Phe Leu 690 695 700 Trp Thr Glu Gin Ala His Thr Val Thr Val Leu Ala Ile Asn Ser Ile 705 710 715 720 0 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 Val Ile Val Ser Trp Ile Leu Ser Pro Ser Asp Tyr Lys Leu Met 755 760 765 Tyr Phe Ile Ile Glu Trp Lys Asn Leu Asn Glu Asp Gly Glu Ile Lys 770 775 780 Trp Leu Arg Ile Ser Ser Ser Val Lys Lys Tyr Tyr Ile His Asp His 785 790 795 800 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 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 i0 Cys Ser Trp Ala Gin Gly Leu Asn Phe Gin Lys Pro Glu Thr Phe Glu 885 890 895 117 His Leu Phe Ile Lys His Thr Ala Ser Val Thr Cys Gly Pro Leu Leu 900 905 910 Leu Glu Pro Glu Thr Ile Ser Glu Asp Ile Ser Val Asp Thr Ser Trp 915 920 925 Lys Asn Lys Asp Glu Met Met Pro Thr Thr Val Val Ser Leu Leu Ser 930 935 940 Thr Thr Asp Leu Glu Lys Gly Ser Val Cys Ile Ser Asp Gin Phe Asn 945 950 955 960 Ser Val Asn Phe Ser Glu Ala Glu Gly Thr Glu Val Thr Tyr Glu Asp 965 970 975 Glu Ser Gin Arg Gin Pro Phe Val Lys Tyr Ala Thr Leu Ile Ser Asn 980 985 990 Ser Lys Pro Ser Glu Thr Gly Glu Glu Gln Gly Leu Ile Asn Ser Ser 995 1000 1005 Val Thr Lys Cys Phe Ser Ser Lys Asn Ser Pro Leu Lys Asp Ser Phe 1010 1015 1020 Ser Asn Ser Ser Trp Glu Ile Glu 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 Glu Gly 1045 1050 1055 Leu Asp Glu Leu Leu Lys Leu Glu Gly Asn Phe Pro Glu Glu Asn Asn 1060 1065 1070 Asp Lys Lys Ser Ile Tyr Tyr Leu Gly Val Thr Ser Ile Lys Lys Arg 1075 1080 1085 Glu Ser Gly Val Leu Leu Thr Asp Lys Ser Arg Val Ser Cys Pro Phe 1090 1095 1100 Pro Ala Pro Cys Leu Phe Thr Asp Ile Arg Val Leu Gin Asp Ser Cys 1105 1110 1115 1120 Ser His Phe Val Glu Asn Asn Ile Asn Leu Gly Thr Ser Ser Lys Lys 45 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 Glu 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 Azg Lys Glu Thr Arg Val Lys Phe Glu Asn Asri 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: SEQUENCE CHARACTERISTICS: LENGTH: 3599 base pairs TYPE: nucleic acid STRANDEDNESS: single TOPOLOGY: linear (ii) MOLECULE TYPE: cDNA (xi) SEQUENCE DESCRIPTION: SEQ ID NO:8: GCGGCCGCCA GTGTGATGGA TATCTGCAGA ATTCGGCTTT GACCCCCGGA TTTTGTTACA o CTCCTTGGAG TGCCTGCTGG AACCTAAGTT GTTGCTTTCG AGACATTTGT o TACAGTGCTG AGAATCTATT TAGAAGATTC GTGTTCACGA TCCTTATGTG o TTCAGCCCAT TCA.AGGTGTA TTGGGA.ATTT ATTTAAGTTG GCTCTCAAAG TAATTCAAGT GAGTGAGCAA TTCAACAGTA GCTAAAAGGA CAGGAATTAT ACCTCTGGTT ATdTTGTGAA TTTGAAAATC AAATATGGTG CTTCTCTGAA ATTTATGTGA TCTTGCATGC AATACTTCAA GGTACTCACT GATAGAAACT AATTCTTTAG GACTTAAAAT AACTATAAGG CCCCAAAA.AG 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 TACT TCCTTT ACAGCTGTTG ACTTTCCACT ATTGAAGGAA AACTGGAACA TCATTATTTA CCTGAAGTGT TGCAATTGCA AACGACACTC CTAATGTCAG GAAATCACAG 120 180 240 300 360 420 480 540 600 660 720 780 840 ATGATGGTAA TTTAAAGATT TCTTGGTCCA GCCCACCATT GGTACCATTT CCACTTCA.AT 119 ATCAAGTGAA ATATTCAGAG CAGCTACATC CCTGCTAGTA GGGGCAAGAG ACTGGATGGc CCACACAAGA TGTCATATAC CTTTTCACTG CATCTATAAG 0 GGATGAATTT AGCTGAGA. GCAAAGTTAC TTTTTTCAAT CAGTGTACTG CTGCAATGA.A ATGTCA.ATAT CAATATCTCA GGTCAACCAG TACAATCCAG .0 GCAGCCTTTA CTGTTCTGAT ATTTGCAGAG TGATGGTTTT 1 ACACAATGTG GATTAGGATC TCCTTCCTGA TTCTGTGGTGA TAAACATTGG ATTATTGAAA A AATTCCAGAT TCGCTATGGT T ATGATGCAAA ATCAAAATCT G AATTCTACA GACAGTATA CCAGGAATC TTTCCACCT. AAGGAAAAC. ATTCCTCAJ CTGAATGAAJ CATGAATGCC rGTGAAACTG TCACTTGCGG kTTCCATCTA ~ATGAATGCA ATCACTCTC AGCCACTGC TATCTTGGG TAAGTGGAA TCAGTCTCC A CAGTTATCAG AGAAGCTGAC C TTCCTGGGTC TTCGTATGAG T GGAGTGACTG GAGTACTCCT A. AAATTCTGAC AAGTGTTGGG A. AGATTGTTCC CTCAAAAGAG SGCCAGTATGA TGTTGTGAGT ~CCAAACCTCG AGGAAAGTTT ATCATCGCTA TGCTGAATTA ATGGGTACTT AACTAAAATG AAAGCACTTT GCAATTGAGG TTCATCCCAT ATCTGAGCCC TTTTCCAGCC AATCTTCCTA TI TAGGTTCACT TGACTCTCCA C CTCCATCCAG TGTGAAAGCA G AAAAGCCAGT CTTTCCAGAG A AAGAAGTACA ATGGA.AGATG T. CAGTTCCAGA CTTGTGTGCA G~ XAGATTGTCT GTTCAGGTGA CGTGTCTTTA TCTAATGTTT ATTGTTTGGT GATCATGTTA ACCTATGATG TATGTGATTG kCTTGCAGAT E'ATCATAGGA LAAGATTGCT 'TATCTGGCT :CAACATGTG AAATTACTA ATAACCTTC P.TGAGGTTT 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 TTCAGGTGCG CTGTAAGAGG CTAGATGGAC TGGGATATTG GAGTAATTGG AGCAATCCAG CCTACACAGT TGTCATGGAT TTAATGGAGA TACTATGA;AA AAAATGACTC ATTGTGCAGT GAACATGGTC AGAAGATGTG CACATACTGT TACGGTTCTG TAACCTTT TC ATGGCCTATG )o TAAACAGCAG TTGTGTGATT ATTTTATTAT TGAGTGGAAA ATAAAAGTTC AAGGAGAAAA GTTCAGAGAT GGAAATCACA GCCATCAATT AGCAAAGTAA GTTTCCTGGA AATCTTAATG CTATGAGAGG ATGTCACTTT ATGTGATAAA CGAAATTCAC CAATTGGTGC ATATCGTGCA TACTATCACC AAGATGGTGA ACCTGAATTT ACTTTGGAAG CCATCATACT TTTCCTGTGG TTCTGTTGCA GTCACTCAGT CAGTGATTAC TGGAGALATAA CCCCTGATGA TCCTGCAATG ACAGAGCAAG A.ATTTTAATT GCTTATCCTT AAGCTAATGT CTTCATCTGT TAAGAAGTAT TATATCCATG ATCATTTTAT CCCCATTGAG AAGTACCAGT 2520 120 TCAGTCTTTA CTCAAGATGA TTATTTCCTC AGCTATTTTG 0 TTCAGAAGCC GTCCTCTTCT AAAATAAAGA AAAAGGGTTC GTACTGAGGT 0 TGATCAGCAA TCACCAAGTG GGGAGATAGA CACACCTCAC AAGAAAATAA ;0 AGAGTGGTGT TATTCACGGA ACTTAGGAAC CCCAATATTT TATTGAAAA-A TTCCATCTTA GGAAGATGTT AGAAACGTTT TTTGGAGCCT TGAGATGATG TGTTTGTATT AACCTATGAG CTCTAAACCA CTTCTCTAGC GGCCCAGGCA ATTCTCAGAA TGATAAAAAG GCTTTTGACT CATCAGAGTT TTCTAGTAAG ATGGAAGGAG CACCAGAGTG TTGCTTGGAA CCGAkACCCCA GAGCATCTTT GAAACAATTT CCAACAACTG AGTGACCAGT GACGAAAGCC AGTGAAACTG AAAAATTCTC TTTTTTATAT GGATTGGATG TCTATCTATT GACAAGTCAA CTCCAGGACA AAGACTTTTG ATGGAAAACA TGGGAAAACC ATGCAGGTTT CATTATTA.AT AGAATTGTTC TTATCAAGCA CAGAAGATAT TGGTCTCTCT TCAACAGTGT AGAGACAACC GTGAAGAACA CGTTGAAGGA TATCGGATCA AACTTTTGAA ATTTAGGGGT GGGTATCGTG GTTGCTCACA CATCTTACAT AGATGTGTGA AAAGATA.ATT ATATGTAATT ATCACACCAA CTGGGCACA.A TACAGCATCA CAGTGTTGAT ACTTTCAACA TA.ACTTCTCT CTTTGTTAA.A AGGGCTTATA TTCTTTCTCT GCATCCCAAC ATTGGAGGGA CACCTCAATC CCCATTCCCA CTTTGTAGAA GCCTCAATTC CCTAACTGTG AATAGTTTCA GTGCCAGTAA AGAATGAAA GGACTTA.ATI' GTGACATGTG ACATCATGGA ACAGATCTTG GAGGCTGAGG TACGCCACGC AATAGTTCAG AATAGCTCAT ATAATTTCAC AATTTCCCTG AAAAAGAGAG GCCCCCTGTT AATAATATCA CAAACTTGTT TAATCTAGA 2580 2640 2700 2760 2820 2880 2940 3000 3060 3120 3180 3240 3300 3360 3420 3480 3540 3599 CTACTCAGAC TCATAAGATC 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 INFORMATION FOR SEQ ID NO:J.0: 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 Gin Lys Phe Cys Val Val Leu Leu His Tyr Phe Leu Giu Asn Arg Ser Ile Giu 145 ValI Lys Pro Thr Leu Asn Thr Gin 130 Ser Ile Leu Ala Ala Ser Cys Val1 115 Cys Leu Thr Ser Gly ValI Lys Ser 100 Asn T rp, Phe Al a ,Cys Leu Giu Thr Leu Ser Leu Lys P he Met Ser Pro 70 Th r Cys Leu Lys As n 150 As n Pro Lys Lys P he Ala Val Gly 135 Leu Leu Pro 40 As n Phe His Asp Phe 120 Asp Phe Ser 25 As n Th r As n Cys Asn 105 Gin Leu A.rg 10 Tyr Ser Se r Ser Cys 90 Ile Gin Lys Asn *Pro Thr As n Ser 75 Phe Giu Ile Leu Tyr 155 Ile Tyr Ser Gly Arg Gly Asp P he 140 As n Trp Thr Asp As n Thr Ser Lys Ala 125 Ile Tyr Giu Pro Tyr Gly His Glu Th r 110 As n Lys Phe ITrp P he His Phe Gin Phe Trp Tyr Val Ile Arg Leu Tyr Ser Asp Val1 As n Val1 His Leu Leu Tyr Vai Pro Giu Val Leu Giu 170 Asp Ser Pro Leu Val Pro 175 122 Gin Lys Gly Ser Phe Gin Met Vai His Cys Asn Cys Ser Val His Giu 180 185 190 Cys Cys Giu Cys Leu Val Pro Val Pro Thr Aia 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 _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 Aia Asp Lys Ile Vai ?0 275 280 285 Ser Aia Thr Ser Leu Leu Vai Asp Ser Ile Leu Pro Giy Ser Ser Tyr 290 295 300 Giu Vai Gin Val Arg Gly Lys Arg Leu Asp Giy Pro Gly Ile Trp Ser 305 310 315 320 Asp Trp Ser Thr Pro Arg Val 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 Ile Tyr Lys Lys Glu Asn Lys Ile Vai Pro Ser Lys Giu Ile Vai Trp 355 360 365 Trp Met Asn Leu Ala Giu Lys Ile Pro Gin Ser Gin Tyr Asp Vai Vai 370 375 380 Ser Asp His Val 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 Leu Tyr Vai 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 Glu Ser *Thr Leu Gin Leu 450 455 460 123 Arg Tyr His Arg Ser 465 Pro Glu Ile Val1 Al.a 545 Pro Ser Ser Val1 T rp 625 Arg Giu Leu Gly T rp 705 Gly Ile Cys Arg Le u 530 Glu Val. Gly Lys Gin 610 Ser Gly Lys Cys Thr 690 Thr Ala Ser Ile Ile Pro Ile Phe Lys Ser 595 Vali As n Pro Asn Ser 675 Trp Glu Ser Pro 485 Gin His Ser Ile Giu 565 Val. Ser Cys Aia Phe 645 Thr Gin Giu Ala Ala 725 Ser Leu 470 Lys Asp Pro Ile Ser Leu Val Val 535 Asn Ile 550 Asn Asn Gin Trp Leu Pro Lys Arg Tyr Thr 630 Trp Arg Leu Leu Arg Tyr Asp Va]. 695 His Thr 710 Asn Phe Ty r Cys Phe Giy 520 Lys Gly Le u Lys Val1 600 Leu Vali Ile T rp Val1 680 Giy Val1 Asn Cys Ser Asp le Pro Ser Ile His Ty r Leu 505 Ser Pro Le u Gin Met 585 Pro Asp Val1 Ile Lys 665 Ile Asn Thr Leu Leu 490 Leu Leu Le u Leu Phe 570 Tyr Asp Giy Met Asn 650 Pro Asn His Val Thr 730 475 Gin Se r Asp Pro Lys 555 Gin Giu Leu Leu Asp 635 Gly Leu His Th r Leu 715 Phe Se r Gly Se r Pro 540 Ile le Vali Cys Gly 620 Ile Asp Met His Lys 700 Ala Ser Asp Tyr Pro 525 Se r Se r Arg Tyr Ala 605 Tyr Lys Th r Lys Thr 685 Phe Ile Trp Giy Th r 510 Pro Se r Trp Ty r Asp 590 Vali T rp Vali Met Asn 670 Ser Th r Asn Pro P he 495 Met Thr Va]. Giu Gly 575 Ala Tyr Ser Pro Lys 655 Asp Cys Phe Ser Met 735 Tyr T rp Cys Lys Lys 560 Le u Lys Ala As n Met 640 Lys Se r Asn Leu Ile 720 Ser 124 Lys Val Asn Cys Val Ile 7~55 Tyr Phe Ile Ile 740 Val Val Gin Ser Leu Ser 745 Se r Ala Tyr Pro Leu Ser Tr Ile Pro Ser Asp Tyr 765 Gly Asn Ser Ser 750 Lys Leu Met Giu Ile Lys Ile Giu Trp 770 Trp Leu Lys 775 Ser Asn Leu Asn Giu Asp 780 Tyr Ile His Asp Arg Ile Ser 785 P he Ser 790 Ly s Vai Lys Lys Tyr 795 Leu His 800 Ile Pro Ile Giu 805 Lys Tyr Gin Phe Ser 810 As n Tyr Pro Ile Phe Met 815 Asp Asp Giu Giy Vai Gly 820 Pro Lys Ile Ser Phe Thr Gin 830 Ile Giu Lys His Gin Ser Asp 835 INFORMIATION FOR SEQ ID NO:ii: SEQUENCE CHARACTERISTICS: LENGTH: 2624 base pairs TYPE: nucieic acid STRANDEDNESS: single TOPOLOGY: linear (ii) MOLECULE TYPE: cDNA (xi) SEQUENCE DESCRIPTION: SEQ ID NO:ii: GCGGCCGCCA GTGTGATGGA TATCTGCAGA ATTCGGCTTT CTCTGCCTTC GACCCCCGGA TCAAGGTGTA CTTCTCTGAA GTAAGATGAT TTGTCAAAAA TTTTGTTACA TTGGGAATTT ATTTATGTGA TAACTGCGTT TAACTTGTCA CTCCTTGGAG ATTTAAGTTG TCTTGCATGC CACCAAATTC AACCTATGAC TGCCTGCTGG GCTCTCAAAG AATACTTCAA ATTCGAATGG ACATTATGAG AACCTAAGTT TAATTCAAGT GGTACTCACT TTTCTAACTT ATCCAAAACA GTTGCTTTCG GAGTGAGCAA GATAGAAACT GCTCCTTATG TGCAGACAAC AGACATTTGT TTCAACAGTA AATTCTTTAG TTTTTCAACA AATAGATGCA TACAGTGCTG GCTAAAAGGA GACTTAAAAT TATTCATCTG TTATGTGGAG GGTCGAGTTG TTCTGTGTGG TATCCAATTA TACTTCCTTT ACAGCTGTTG ACTTTCCACT ATTGAAGGAA AACTGGAACA TCATTATTTA 120 180 240 300 360 420 480 540 125 AGAATCTATT CAGGAATTAT AACTATAAGG TCCATCTTTT ATATGTTCTG CCTGA.AGTGT TAGAAGATTC ACCTCTGGTT CCCCAAAAAG GCAGTTTTCA GATGGTTCAC TGCAATTGCA GTGTTCACGA TCCTTATGTG TTCAGCCCAT ATGATGGTAA ATGTTGTGAA TTTGAAA.ATC AA.ATATGGTG TTTAAAGATT TGTCTTGTGC ACATCTGGTG AAGCCTGATC TCTTGGTCCA AATTCTACAA CTGTGCCAAC AGCCAAACTC A-ACGACACTC ATCAAGTGA. ATATTCAGAG CAGCTACATC GGGGCAAGAG J CCACACAAGA CTTTTCACTG GGATGAATTT GCAAAGTTAC CAGTGTACTG o ATGTCAATAT GGTCAACCAG GCAGCCTTTA ATTTGCAGAG ACACAATGTG o TCCTTCCTGA TAAACATTGG AATTCCAGAT ATGATGCAAA Iz TTCAGGTGCG C o CCTACACAGT T TTAATGGAGA TI CCTGCTAGTA GACAGTATA ACTGGA'IGGC CCAGGAATC TGTCATATAC TTTCCACCT CATCTATAAG A.AGGAAAAC. AGCTGAGA ATTCCTCAA, TTTTTTCAAT CTGAATGAAj CTGCAATGA CATGAATGC( CAATATCTCA TGTGAAACTC TACAATCCAG TCACTTGCGC CTGTTCTGAT ATTCCATCTA TGATGGTTTT TATGAATGCA GATTAGGATC AATCACTCTC TCTGTGGTG AAGCCACTGC kTTATTGAAA ATATCTTGGG C'CGCTATGGT TTAAGTGGAA ~TCAAAATCT GTCAGTCTCC .TGTAAGAGG CTAGATGGAC 'GTCATGGAT ATAAA.AGTTC ACTATGA. AAGGAGAAAA 'C T A A A C' GAGTAATTTT CACCATTAGG GCCCACCATT CAGTTATCAG TTCCTGGGTC GGAGTGACTG AAATTCTGAC AGATTGTTCC GCCAGTATGA CCAAACCTCG k.TCATCGCTA k.TGGGTACTT LAAGCACTTT ~TCATCCCAT 'TTTCCAGCC 'AGGTTCACT I1 TCCATCCAG 'T AAAGCCAGT C AGAAGTACA A PiGTTCCAGA C CCAGTCACCT TTTGCATATG GGTACCATTT AGAAGCTGAC TTCGTATGAG GAGTACTCCT A.AGTGTTGGG CTCAAAAGAG TGTTGTGAGT AGGAAAGTTT TGCTGAATTA ;LACTAAAATG GCAATTGAGG kTCTGAGCCC LkTCTTCCTA I~ 'GACTCTCCA C ~GTGAAAGCA G :TTTCCAGAGA .TGGAAGATG T TTGTGTGCA G CTAATGTCAG GA.AATCACAG CCACTTCA-AT AAGATTGTCT GTTCAGGTGA CGTGTCTTTA TCTAATGTTT ATTGTTTGGT GATCATGTTA ACCTATGATG rATGTGATTG kCTTGCAGAT rATCATAGGA LkAGATTGCT ~TATCTGGCT CAACATGTG AAATTACTA ATAACCTTC ATGAGGTTT TCTATGCTG GCAATCCAG GGAGAATAA CCCTGATGA 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 GGGATATTG GAGTAATTGG CTATGAGAGG ATGTCACTTT ACCTGA.ATTT ACTTTGGAAG 126 AAAATGACTC ATTGTGCAGT GTTCAGAGAT ATGTGATAAA GAACA*TGGTC AGA.AGATGTG GGAAATCACA CGAAATTCAC CACATACTGT TACGGTTCTG GCCATCAATT CAATTGGTGC TAACCTTTTC ATGGCCTATG AGCAA.AGTAA ATATCGTGCA TAAACAGCAG TTGTGTGATT GTTTCCTGGA TACTATCACC 0 ATTTTATTAT TGAGTGGAAA AATCTTAATG AAGATGGTGA CTTCATCTGT TAkAGA.AGTAT TATATCCATG ATCATTTTAT TCAGTCTTTA CCCAATATTT ATGGAAGGAG TGGGAAAACC CTCAAGATGA TATTGAAAAA CACCAGAGTG ATTGATAAGG INFORMATION FOR SEQ ID NO:l2: Ci) SEQUENCE CHARACTERISTICS: LENGTH: 2948 base pairs TYPE: nucleic acid STRANDEDNESS: single TOPOLOGY: linear (ii) MOLECULE TYPE: cDNA CCATCATACT TTTCCTGTGG TTCTGTTGCA GTCACTCAGT CAGTGATT'AC AATAAAATGG CCCCATTGAG AAAGATAATT ATCC TCCTGCAATG ACAGAGCAAG AATTTTAAT T GCTTATCCTT AAGCTAATGT CTTAGAATCT AAGTACCAGT AATAGTTTCA 2160 2220 2280 2340 2400 2460 2520 2580 2624 (xi) SEQUENCE DESCRIPTION: SEQ ID.NO:12: CCATTGAAGT CAATGGGAGT TTGTTTTGGC ACCAAAATCA ACGGGGATTT CCAAAATGTC GTAATAACCC TAAGCAGAGC AGCTTGCTAG GTCGAGTTGG TCTGTGTGGT ATCCAATTAC ACTTCCTTTT CAGCTGTTGA CTTTCCACTG CGCCCCGTTG TCGTTTAGTG CGGCCGCCAG ACCCCCGGAT TTTGTTACAT TCCTTGGAGA GCCTGCTGGG ACCTAAGTTT TTGCTTTCGG ACGCAAATGG AACCGTCAGA TGTGATGGAT CAAGGTGTAC TGGGAATTTA TTTAAGTTGT CTCTCAAAGA AATTCAAGTG AGTGAGCAAG GCGGTAGGCG TCTCTAGAAG ATCTGCAGALA TTCTCTGAAG TTTATGTGAT CTTGCATGCC ATACTTCAAA GTACTCACTT ATAGAAACTG TGTACGGTGG CTGGGTACCA TTCGGCTTTC TX.AGATGATT AACTGCGTTT ACCAAATTCA TTCGAATGGA TTCTAACTTA CTCCTTATGT GAGGTCTATA GCTGCTAGCA TCTGCCTTCG TGTCAAAAAT AACTTGTCAT ACCTATGACT CATTATGAGA TCCAAAACAA GCAGACAACA 127 TTGAAGGAAA ACTGGA.ACAT CATTATTTA.A CTGAAGTGTT GCAATTGCAG ACGACACTCT GACATTTGTT ACAGTGCTGG GAATCTATTC AGAAGATTCA TGTTCACGAA CCTTATGTGT TAATGTCAGT TCAGCCCATA *AAATCACAGA CACTTCAATA AGATTGTCTC TTCAGGTGAG GTGTCTTTAC CTAATGTTTC TTGTTTGGTG ATCATGTTAG CCTATGATGC ATGTGATTGA 3 CTTGCAGATG ATCATAGGAG AAGATTGCTA TATCTGGCTA CAACATGTGT AAATTACTAT ATAACCTTCA ATGAGGTTTA TCTATGCTGT TGATGGTAAT TCAAGTGAA.A AGCTACATCC GGGCAAGAGA CACACAAGAT TTTTCACTGC GATGAATTTA CAAAGTTACT AGTGTACTGC TGTCAATATC GTCAACCAGT CAGCCTTTAC TTTGCAGAGT CACAATGTGG CCTTCCTGAT AAACATTGGA ATTCCAGATT TGATGCAAAA TCAGGTGCGC TCA.ACAGTAA CTAAAAGGAG AGGAATTATA CCTCTGGTTC TGTTGTGA.AT TTGA.AAATCA AATATGGTGA TTAAAGATTT TATTCAGAGA CTGCTAGTAG CTGGATGGCC GTCATATACT ATCTATAAGA GCTGAGAAAA TTTTTCAATC TGCAATGAAC AATATCTCAT ACAATCCAGT TGTTCTGATA GATGGTTTTT ATTAGGATCA TCTGTGGTGA TTATTGAAAA CGCTATGGTT TCAAAATCTG TGTAAGAGGC ATTCTTTAG-T ACTTAAAATT ACTATAAGGT CCCAAAA.AGG GTCTTGTGCC CATCTGGTGG AGCCTGATCC CTTGGTCCAG ATTCTACAAC ACAGTATACT CAGGAATCTG TTCCACCTAA AGGAAAACAA TTCCTCAAAG- TGA.ATGAAAC ATGAATGCCA GTGAAACTGA CACTTGCGGA TTCCATCTAT ATGAATGCAT ATCACTCTCT AGCCACTGCC TATCTTGGGA TAAGTGGAAA TCAGTCTCCC TAGATGGACT TT-TTCA.ACAA ATTCATCTGT CCATCTTTTA CAGTTTTCAG TGTGCCAACA AGTAATTTTC ACCATTAGGT CCCACCATTG AGTTATCAGA TCCTGGGTCT GAGTGACTGG KATTCTGACA GATTGTTCCC CCAGTATGAT CAA.ACCTCGA TCATCGCTAT TGGGTACTTA A-AGCACTTTG TCATCCCATA TTTCCAGCCA AGGTTCACTT TCCATCCAGT AAAGCCAGTC AGAAGTACAA AGTTCCAGAC GGGATATTGG ATAGATGCAA TATGTGGAGT TATGTTCTGC ATGGTTCACT GCCAAACTCA CAGTCACCTC TTGCATATGG GTACCATTTC GAAGCTGACA TCGTATGAGG AGTACTCCTC AGTGTTGGGT TCAAAAGAGA GTTGTGAGTG GGAAAGTTTA GCTGAATTAT ACTAAAATGA CA.ATTGAGGT TCTGAGCCCA ATCTTCCTAT GACTCTCCAC GTGAAAGCAG TTTCCAGAGA TGGAAGATGT TTGTGTGCAG AGTAATTGGA 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 GCAATCCAGC CTACACAGTT GTCATGGATA TAAAAGTTCC TATGAGAGGA CCTGAATTTT 128 GGAGAATAAT CCCTGATGA.A CCTGCAATGG CAGAGCAAGC ATTTTA.ATTT CTTATCCTTT AGCTAATGTA TTAGAATCTC AGTACCAGTT JATAGTTTCAC ACGATGACAA ACTAGAATGC GTAACCAT TAATGGAGAT AAATGACTCA AACATGGTCA ACATACTGTT AACCTTTTCA AAACAGCAGT TTTTATTATT TTCATCTGTT CAGTCTTTAC TCAAGATGAT GTAGGGATCC AGTGAAAAAA ACTATGAAA TTGTGCAGTG GA.AGATGTGG ACGGTTCTGG TGGCCTATGA TGTGTGATTG GAGTGGAAAA AAGAAGTATT CCAATATTTA ATTGAAAAAC AGACATGATA ATGCTTTATT AGGAGAAAA-A TGTCACTTTA CTTTGGAAGC TTCAGAGATA GAAATCACAC CCATCAATTC GCAAAGTAAA TTTCCTGGAT ATCTTAATGA ATATCCATGA TGGAAGGAGT ACCAGAGTGA AGATACATTG TGTGAAATTT TGTGATAAAC GAAATTCACT AATTGGTGCT TATCGTGCAG ACTATCACCC AGATGGTGAA TCATTTTATC GGGAAAACCA TGCAGGTGAC ATGAGTTTGG GTGATGCTAT CATCATACTT TTCCTGTGGA TCTGTTGCA.A TCACTCAGTG AGTGATTACA ATAAAATGGC CCCATTGAGA AAGATAATTA TACAAGGACG ACAACCCACA TGCTTTATTT 2280 2340 2400 2460 2520 2580 2640 2700 2760 2820 2880 2940 2948 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:13: Met Ile Cys Gin Lys Phe Cys Val Val Leu 1 5 10 Tyr Val Ile Thr Ala Phe Asn Leu Ser Tyr 25 Phe Lys Leu Ser Cys Met Pro Pro Asn Ser 40 Leu Pro Ala Gly Leu Ser Lys Asn Thr Ser s0 55 Leu His Trp Glu Phe Ile Pro Ile Thr Pro Trp Arg Thr Tyr Asp Tyr Phe Leu Asn Ser Asn Gly His Tyr 129 Giu Thr Ala Asn Leu Ser Arg Asn Cys Ser Thr Val 115 Ile Gin Cys 130 Giu Ser Leu 145 Leu Leu Tyr Gin Lys Gly Cys Cys Glu 195 Leu Leu Met 210 Pro Leu Met 225 Leu Gly Leu Trp Ser Ser Tyr Ser Glu 275 Ser Ala Thr 290 Giu Val Gin 305 Val Giu Lys Thr Ser Leu 100 Asn Ser Trp Leu Phe Lys Val Leu 165 Ser Phe 180 Cys Leu Cys Leu Ser Vai His Met 245 Pro Pro 260 Asn Ser Ser Leu Val Arg Pro Lys Thr Phe Cys Ala Leu Val Lys Gly 135 Asn Leu 150 Pro Giu Gin Met Val Pro Lys Ile 215 Gin Pro 230 Giu Ile Leu Val Thr Thr Leu Val 295 Gly Lys 310 P he His Asp P he 120 Asp P he Val1 Val1 Val1 200 Th r Ile Th r Pro Val 280 Asp Arg As n Cys As n 105 Gin Leu Arg Leu His 185 Pro Ser As n Asp Phe 265 Ile Ser Leu Ser Ser Giy Thr His Phe Ser 75 Cys 90 Ile Gin Lys As n Giu 170 Cys Thr Gly Met Asp 250 Pro Arg Ile Asp Phe Giu Ile Leu Ty r 155 Asp As n Al a Gly Val 235 Gly Leu Giu Leu Gly 315 Azg Gly Asp P he 140 As n Ser Cys Lys Val1 220 Lys As n Gin Ala Pro 300 Pro Ser Lys Al a 125 Ile Tyr Pro Se r Leu 205 Ile Pro Leu Tyr Asp 285 Gly Gly Glu Thr 110 As n Cys Lys Leu ValI 190 As n Phe Asp Lys Gin 270 Lys Ser Ile Gin Phe T rp Tyr Val1 Val1 175 His Asp Gin Pro Ile 255 Val1 Ile Ser Trp Asp Val As n Val1 His 160 Pro Giu Thr Ser Pro 240 Ser Lys Val1 Tyr Ser 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 340 Ile Trp Ser 385 Pro Glu Asn Trp Arg 465 Pro Glu Ile Val Ala 545 Pro Ser Ser Tyr Met 370 Asp Arg Cys Ile Ser 450 Tyr Ile Cys Arg Leu 530 Glu Val Gly Lys Lys 355 Asn His Gly His Ser 435 Thr His Ser Ile Ile 515 Pro Ile Phe Lys Ser 595 Lys Leu Val Lys His 420 Cys Ser Arg Glu Phe 500 Asn Asp Thr Pro Glu 580 Val Glu Ala Ser Phe 405 Arg Glu Thr Ser Pro 485 Gin His Ser Ile Glu 565 Val Ser Asn Glu Lys 390 Thr Tyr Thr Ile Ser 470 Lys Pro Ser Val Asn 550 Asn Gin Leu 130 Ser Val Lys lle 360 Lys Ile 375 Val Thr Tyr Asp Ala Glu Asp Gly 440 Gin Ser 455 Leu Tyr Asp Cys Ile Phe Leu Gly 520 Val Lys 535 Ile Gly Asn Leu Trp Lys Pro Val 600 Gly 345 Val Pro Phe Ala Leu 425 Tyr Leu Cys Tyr Leu 505 Ser Pro Leu Gin Met 585 Pro Ser Pro Gin Phe Val 410 Tyr Leu Ala Ser Leu 490 Leu Leu Leu Leu Phe 570 Tyr Asp Asn SSer Ser Asn 395 Tyr Val Thr Glu Asp 475 Gin Ser Asp Pro Lys 555 Gin Glu Leu Val Lys Gin 380 Leu Cys le Lys Ser 460 Ile Ser Gly Ser Pro 540 Ile Ile Val Cys Ser Glu 365 Tyr Asn Cys Asp Met 445 Thr Pro Asp Tyr Pro 525 Ser Ser Arg Tyr Ala 605 Phe 350 Ile Asp Glu Asn Val 430 Thr Leu Ser Gly Thr 510 Pro Ser Trp Tyr Asp 590 Val His Val Val Thr Glu 415 Asn Cys Gin Ile Phe 495 Met Thr Val Glu Gly 575 Ala Tyr Cys Trp Val Lys 400 His Ile Arg Leu His 480 Tyr Trp Cys Lys Lys 560 Leu Lys Ala Val Gin Val Arg Cys Lys Arg Leu Asp Gly Leu Gly Tyr Trp Ser Asn 131 Trp Ser Asn Pro Ala Tyr Thr V~ 625 630 Arg Gly Pro Giu Phe Trp Arg 11 645 Giu Lys Asn Val Thr Leu Leu Tr 660 Leu Cys Ser Vai Gin Arg Tyr Va 675 68 Giy Thr Trp Ser Giu Asp Val Gil 690 695 Trp Thr Glu Gin Ala His Thr Val 705 710 Gly Ala Ser Val Ala Asn Phe Asn 725 Lys Val Asn Ile Val Gin Ser Leu 7.40 Cys Val Ile Vai Ser Trp Ile Leu 755 760 Tyr Phe Ile Ile Giu Trp Lys Asn 770 775 Trp Leu Arg Ile Ser Ser Ser Val 785 790 Phe Thr Ile Leu INFORMATION FOR SEQ ID NO:14: Ci) SEQUENCE CHARACTERISTICS: LENGTH: 2507 base pairs TYPE: nucleic acid STRANDEDNESS: single TOPOLOGY: linear MOLECULE TYPE: cDNA 1 Val met e Ile Asn 650 p Lys pro 665 L Ile Asn Asn His Thr Val Leu Thr P 730 Ser Ala TI 745 Ser Pro S Leu Asn G. Lys Lys T3 GI Le Hi. Th Leu ~he yr er lu p Ilie y Asp u Met a His rLys 700 Ala Ser 'I Pro L. Asp T 7 Asp G. 780 Tyr I2 Ly Ly Thi 685 P he I le rp e u yr J-y Le r me 3 Asj 67~ Se: Thr Asn Pro As n '750 Lys Glu His I Pr t Ly 65 nl As rCy~ Ph Se r Met 735 Ser Le u Ile Gly 0 Met 640 5S Lys p Ser S Asn SLeu Ile 720 Se r Se r Met Lys Lys 800 132 (xi) SEQUENCE DESCRIPTION: SEQ ID NO:14: GCGGCCGCCA GTGTGATGGA TATCTGCAGA ATTCGGCTTT CTCTGCCTTC GGTCGAGTTG GACCCCCGGA TTTTGTTACA CTCCTTGGAG 0 TGCCTGCTGG AACCTAAGTT GTTGCTTTCG AGACATTTGT TACAGTGCTG 0 AGAATCTATT TAGAAGATTC TCAAGGTGTA TTGGGAATTT ATTTAAGTTG GCTCTCAA-AG TAATTCAAGT GAGTGAGCAA TTCAACAGTA GCTAAA.AGGA CAGGAATTAT ACCTCTGGTT GTGTTCACGA ATGTTGTGAA TCCTTATGTG TTTGAA.AATC TTCAGCCCAT AAATATGGTG 0 ATGATGGTAA TTTAAAGATT ATCAAGTGAA ATATTCAGAG CAGCTACATC CCTGCTAGTA GGGGCAAGAG ACTGGATGGC CCACACAAGA TGTCATATAC 0 CTTTTCACTG CATCTATAAG GGATGAATTT AGCTGAGAAA GCAAAGTTAC TTTTTTCAAT CAGTGTACTG CTGCAATGAA ATGTCAATAT CAATATCTCA GGTCAACCAG TACAATCCAG CTTCTCTGAA ATTTATGTGA TCTTGCATGC AATACTTCAA GGTACTCACT GATAGAAACT AATTCTTTAG GACTTAAAAT AACTATAAGG CCCCAAAAAG TGTCTTGTGC ACATCTGGTG AAGCCTGATC TCTTGGTCCA AATTCTACAA GACAGTATAC CCAGGAATCT TTTCCACCTA AAGGAAAACA hTTCCTCAAA ZTGAATGAAA ::ATGAATGCC rGTGAAACTG2 rCACTTGCGG2 GTAAGATGAT TAACTGCGTT CACCAAATTC ATTCGAATGG TTTCTA.ACTT GCTCCTTATG TTTTTCA.ACA TATTCATCTG TCCATCTTTT GCAGTTTTCA CTGTGCCAAC GAGTAATTTT CACCATTAGG GCCCACCATT CAGTTATCAG TTCCTGGGTC GGAGTGACTG AAATTCTGAC ALGATTGTTCC GCCAGTATGA :CAA.ACCTCG k.TCATCGCTA kTGGGTACTT LkAGCACTTT TTGTCAA TAACTTGTCA AACCTATGAC ACATTATGAG ATCCAAAACA TGCAGACAAC AATAGATGCA TTATGTGGAG ATATGTTCTG GATGGTTCAC AGCCAAACTC CCAGTCACCT TTTGCATATG GGTACCATTT AGAAGCTGAC TTCGTATGAG GAGTACTCCT AAGTGTTGGG CTCAAAAGAG TGTTGTGAGT AGGAAAGTTT TGCTGAATTA A.ACTAAAATG GCAATTGAGG TTCTGTGTGG TATCCAATTA TACTTCCTTT ACAGCTGTTG ACTTTCCACT ATTGA.AGGAA AACTGGAACA TCATTATTTA CCTGAAGTGT TGCAATTGCA AACGACACTC CTAATGTCAG GAAATCACAG CCACTTCA.AT AAGATTGTCT GTTCAGGTGA CGTGTCTTTA TCTAATGTTT A.TTGTTTGGT GATCATGTTA ACCTATGATG rATGTGATTG ACTTGCAGAT rATCATAGGA 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 GCAGCCTTTA CTGTTCTGAT ATTCCATCTA TTCATCCCAT ATCTGAGCCC AAAGATTGCT 133 ATTTGCAGAG ACACAATGTG TCCTTCCTGA TAAACATTGG J AATTCCAGAT ATGATGCAAA TTCAGGTGCG CCTACACAGT TTAATGGAGA 0 AAAATGACTC GAACATGGTC CACATACTGT TAACCTTTTC TAAACAGCAG o ATTTTATTAT TGATGGTTTT GATTAGGATC TTCTGTGGTG ATTATTGAA.A TCGCTATGGT ATCAAA.ATCT CTGTAAGAGG TGTCATGGAT TACTATGAAA ATTGTGCAGT AGAAGATGTG TACGGTTCTG ATGGCCTATG TTGTGTGATT TGAGTGGAAA TATGAATGCA XATCACTCTC AAGCCACTGC ATATCTTGGG TTAAGTGGAA GTCAGTCTCC CTAGATGGAC ATAAA.AGTTC AAGGAGAAAA GTTCAGAGAT GGAAATCACA GCCATCAATT AGCAAAGTAA GTTTCCTGGA AATCTTAATG TTTTCCAGCC A.ATCTTCCTA TTATCTGGCT TAGGTTCACT CTCCATCCAG AAAAGCCAGT AAGAAGTACA CAGTTCCAGA TGGGATATTG CTATGAGAGG ATGTCACTTT ATGTGATAAA CGAAATTCAC CAATTGGTGC ATATCGTGCA TACTATCACC AAGATGGTGA GTAAGTTTAC TGACTCTCCA TGTGAAAGCA CTTTCCAGAG ATGGAAGATG CTTGTGTGCA GAGTA.ATTGG ACCTGAATTT ACTTTGGAAG CCATCATACT TTTCCTGTGG TTCTGTTGCA GTCACTCAGT CAGTGATTAC AATAAA.ATGG TATACTT CCALACATGTG GAAATTACTA AkATAACCTTC TATGAGGTTT GTCTATGCTG AGCAATCCAG TGGAGA.ATAA CCCCTGATGA TCCTGCAATG ACAGAGCAAG AATTTTAATT GCTTATCCTT AAGCTA.ATGT CTTAGAATCT 1620 1680 1740 1800 1860 1920 1980 2040 2100 2160 2220 2280 2340 2400 2460 2507 CTTCATCTGT TAAGAAGTAT TATATCCATG INFORMATION FOR SEQ ID 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 29 134 INFORMATION FOR SEQ ID NO:16: SEQUENCE CHARACTERISTICS: LENGTH: 29 base pairs TYPE: nucleic acid STRANDEDNESS: single TOPOLOGY: linear (ii) MOLECULE TYPE: cDNA (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 TOPOLOGY: linear (ii) MOLECULE TYPE: cDNA 0 (xi) SEQUENCE DESCRIPTION: SEQ ID NO:17: GTAAGTACCA AANNNNNTTT TCAATATAG 29 INFORMATION FOR SEQ ID NO:18: SEQUENCE CHARACTERISTICS: LENGTH: 29 base pairs TYPE: nucleic acid 0 STRANDEDNESS: single TOPOLOGY: linear (ii) MOLECULE TYPE: cDNA (xi) SEQUENCE DESCRIPTION: SEQ ID NO:18: 0 GTAAGTTATG CANNNNNTTT TTCCTTAAG oa 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 (ii) MOLECULE TYPE: cDNA ;0 (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: cDNA (xi) SEQUENCE DESCRIPTION: SEQ ID NO:21: GTAAGAAAAC AGNNNNNTGT TTCAAATAG 29 136 INFORMATION FOR SEQ ID NO:22: SEQUENCE CHARACTERISTICS: LENGTH: 29 base pairs S(B) 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 3 (ii) MOLECULE TYPE: cDNA 3 (xi) SEQUENCE DESCRIPTION: SEQ ID NO:23: GTATGTCAAG CTNNNNNAAA AATTTCTAG 29 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 0 (xi) SEQUENCE DESCRIPTION: SEQ ID NO:26: GTATTCCCAA TTNNNNNTAT TTACTACAG 29 INFORMATION FOR SEQ ID NO:27: SEQUENCE CHARACTERISTICS: LENGTH: 29 base pairs 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: 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: S GTATCCAGTG TTNNNNNCTT TTTAAACAG 29