Adipocyte-specific protein homologs

Abstract

The present invention relates to polynucleotide and polypeptide molecules for zsig39, a novel member of the family of proteins bearing a collagen-like domain and a globular domain. The polypeptides, and polynucleotides encoding them, are involved in dimerization or oligomerization and may be used in the study thereof. The present invention also includes antibodies to the zsig39 polypeptides.

What is claimed is:  
     
         1 . An isolated polypeptide comprising a sequence of amino acid residues that is at least 80% identical to SEQ ID NO:2, wherein said sequence comprises: 
 beta strands corresponding to amino acid residues 105-109, 128-130, 136-139, 143-146, 164-171, 176-182, 187-200, 204-210 and 226-231 of SEQ ID NO:2, wherein the beta strands are separated by at least two amino acid residues; and    a receptor binding domain comprising amino acid residues 111-135 and 170-174 of SEQ ID NO:2.    
     
     
         2 . An isolated polypeptide according to  claim 2 , wherein said polypeptide is at least 90% identical to SEQ ID NO:2.  
     
     
         3 . An isolated polypeptide according to  claim 2 , wherein said polypeptide comprises a collagen-like domain having at least 22 collagen repeats.  
     
     
         4 . An isolated polypeptide according to  claim 2 , wherein said polypeptide comprises residues 19-243 of SEQ ID NO:2.  
     
     
         5 . An isolated polypeptide according to  claim 1 , covalently linked amino terminally or carboxy terminally to a moiety selected from the group consisting of affinity tags, toxins, radionucleotides, enzymes and fluorophores.  
     
     
         6 . An isolated polypeptide selected from the group consisting of: 
 a) a polypeptide having a sequence of amino acid residues from amino acid residue 30 to amino acid residue 95 of SEQ ID NO:2;    b) a polypeptide having a sequence of amino acid residues from amino acid residue 30 to amino acid residue 96 of SEQ ID NO:2; and    c) a polypeptide having a sequence of amino acid residues from amino acid residue 30 to 97 of SEQ ID NO:2;    d) a polypeptide having a sequence of amino acid residues from amino acid residue 30 to amino acid residue 98 of SEQ ID NO:2;    e) a polypeptide having a sequence of amino acid residues from amino acid residue 98 to amino acid residue 243 of SEQ ID NO:2;    f) a polypeptide having a sequence of amino acid residues from amino acid residue 99 to amino acid residue 243 of SEQ ID NO:2;    g) a polypeptide having a sequence of amino acid residues from amino acid residue 30 to amino acid residue 243 of SEQ ID NO:2; and    h) a polypeptide having a sequence of amino acid residues that is 90% identical in amino acid sequence to a), b), c), d), e), f), g) or h).    
     
     
         7 . A fusion protein consisting essentially of a first portion and a second portion joined by a peptide bond, said first portion comprising a polypeptide selected from the group consisting of: 
 a) a polypeptide comprising a sequence of amino acid residues that is at least 80% identical to SEQ ID NO:2, wherein said sequence comprises: beta strands corresponding to amino acid residues 105-109, 128-130, 136-139, 143-146, 164-171, 176-182, 187-200, 204-210 and 226-231 of SEQ ID NO:2, wherein the beta strands are separated by at least two amino acid residues; and a receptor binding domain comprising amino acid residues 111-135 and 170-174 of SEQ ID NO:2;    b) a polypeptide comprising a sequence of amino acid residues as shown in SEQ ID NO:2 from amino acid residue 16 to amino acid residue 243;    c) a polypeptide comprising a sequence of amino acid residues as shown in SEQ ID NO:2 from amino acid residue 1 to amino acid residue 243;    d) a portion of the zsig39 polypeptide as shown in SEQ ID NO:2 containing the collagen-like domain or a portion of the collagen-like domain capable of dimerization or oligomerization;    e) a portion of the zsig39 polypeptide as shown in SEQ ID NO:2, containing the globular-like domain or the receptor binding portion of the globular-like domain; or    f) a portion of the zsig39 polypeptide as shown in SEQ ID NO:2, including the collagen-like domain and the globular domain; and    said second portion comprising another polypeptide.    
     
     
         8 . A fusion protein according to  claim 7 , wherein said first portion is selected from the group consisting of: 
 a) a polypeptide having the sequence of amino acid residue 30 to amino acid residue 95 of SEQ ID NO:2;    b) a polypeptide having the sequence of amino acid residue 30 to amino acid residue 96 of SEQ ID NO:2;    c) a polypeptide having the sequence of amino acid residue 30 to amino acid residue 97 of SEQ ID NO:2;    d) a polypeptide having the sequence of amino acid residue 30 to amino acid residue 98 of SEQ ID NO:2;    e) a polypeptide having the sequence of amino acid residue 30 to amino acid residue 243 of SEQ ID NO:2;    f) a polypeptide having the sequence of amino acid residue 98 to amino acid residue 243 of SEQ ID NO:2; and    g) a polypeptide having the sequence of amino acid residue 99 to amino acid residue 243 of SEQ ID NO:2.    
     
     
         9 . A fusion protein comprising a secretory signal sequence having the amino acid sequence of amino acid residues 1-15 or 1-18 of SEQ ID NO:2, wherein said secretory signal sequence is operably linked to an additional polypeptide.  
     
     
         10 . A pharmaceutical composition comprising a polypeptide according to  claim 1 , in combination with a pharmaceutically acceptable vehicle.  
     
     
         11 . An antibody that specifically binds to an epitope of a polypeptide according to  claim 1 .  
     
     
         12 . An isolated polynucleotide encoding a polypeptide comprising a sequence of amino acid residues that is at least 80% identical to SEQ ID NO:2, wherein said sequence comprises: 
 beta strands corresponding to amino acid residues 105-109, 128-130, 136-139, 143-146, 164-171, 176-182, 187-200, 204-210 and 226-231 of SEQ ID NO:2, wherein the beta strands are separated by at least two amino acid residues; and    a receptor binding domain comprising amino acid residues 111-135 and 170-174 of SEQ ID NO:2.    
     
     
         13 . An isolated polynucleotide according to  claim 12 , wherein said polypeptide is at least 90% identical to SEQ ID NO:2.  
     
     
         14 . An isolated polynucleotide according to  claim 12 , wherein said polypeptide comprises a collagen-like domain having at least 22 collagen repeats.  
     
     
         15 . An isolated polynucleotide according to  claim 12 , wherein said polynucleotide is DNA.  
     
     
         16 . An isolated polynucleotide selected from the group consisting of: 
 a) a sequence of nucleotides from nucleotide 243 to nucleotide 962 of SEQ ID NO:1;    b) a sequence of nucleotides from nucleotide 252 to nucleotide 962 of SEQ ID NO:1;    c) a sequence of nucleotides from nucleotide 285 to nucleotide 482 of SEQ ID NO:1;    d) a sequence of nucleotides from nucleotide 285 to nucleotide 485 of SEQ ID NO:1;    e) a sequence of nucleotides from nucleotide 285 to nucleotide 488 of SEQ ID NO:1;    f) a sequence of nucleotides from nucleotide 285 to nucleotide 491 of SEQ ID NO:1;    g) a sequence of nucleotides from nucleotide 285 to nucleotide 926 of SEQ ID NO:1;    h) a sequence of nucleotides from nucleotide 491 to nucleotide 926 of SEQ ID NO:1;    i) a polynucleotide encoding a polypeptide having a sequence of nucleotides that is at least 80% identical in nucleotide sequence to a), b), c), d), e), f), g) and h);    j) nucleotide sequences complementary to a), b), c), d), e), f), g), h) or i); and    k) degenerate nucleotide sequences of a), b), c) d), e), f), g), h), i) or j).    
     
     
         17 . An isolated polynucleotide encoding a fusion protein consisting essentially of a first portion and a second portion joined by a peptide bond, said first portion is selected from the group consisting of: 
 a) a polypeptide comprising a sequence of amino acid residues that is at least 80% identical to SEQ ID NO:2, wherein said sequence comprises: beta strands corresponding to amino acid residues 105-109, 128-130, 136-139, 143-146, 164-171, 176-182, 187-200, 204-210 and 226-231 of SEQ ID NO:2, wherein the beta strands are separated by at least two amino acid residues; and a receptor binding domain comprising amino acid residues 111-135 and 170-174 of SEQ ID NO:2;    b) a polypeptide comprising a sequence of amino acid residues as shown in SEQ ID NO:2 from amino acid residue 16 to amino acid residue 243;    c) a polypeptide comprising a sequence of amino acid residues as shown in SEQ ID NO:2 from amino acid residue 1 to amino acid residue 243;    d) a portion of the zsig39 polypeptide as shown in SEQ ID NO:2 containing the collagen-like domain or a portion of the collagen-like domain capable of dimerization or oligomerization;    e) a portion of the zsig39 polypeptide as shown in SEQ ID NO:2, containing the globular-like domain or an active portion of the globular-like domain; or    f) a portion of the zsig39 polypeptide as shown in SEQ ID NO:2, including the collagen-like domain and the globular domain; and    said second portion comprising another polypeptide.    
     
     
         18 . An isolated polynucleotide encoding a fusion protein comprising a secretory signal sequence having the amino acid sequence of amino acid residues 1-15 or 1-18 of SEQ ID NO:2, wherein said secretory signal sequence is operably linked to an additional polypeptide.  
     
     
         19 . An isolated polynucleotide comprising the sequence of nucleotide 1 to nucleotide 729 of SEQ ID NO:10.  
     
     
         20 . An expression vector comprising the following operably linked elements: 
 a transcription promoter;    a DNA segment encoding a polypeptide according to  claim 1;  and    a transcription terminator.    
     
     
         21 . An expression vector according to  claim 20 , wherein said DNA segment encodes a polypeptide that is at least 90% identical to SEQ ID NO:2.  
     
     
         22 . An expression vector according to  claim 20 , wherein said DNA segment encodes a polypeptide further comprising a collagen-like domain having at least 22 collagen repeats.  
     
     
         23 . An expression vector according to  claim 20 , wherein said DNA segment encodes a polypeptide covalently linked amino terminally or carboxy terminally to an affinity tag.  
     
     
         24 . An expression vector according to  claim 20  wherein said DNA segment further encodes a secretory signal sequence operably linked to said polypeptide.  
     
     
         25 . An expression vector according the  claim 20 , wherein said secretory signal sequence comprises residues 1-15 or 1-18 of SEQ ID NO:2.  
     
     
         26 . A cultured cell into which has been introduced an expression vector comprising the following operably linked elements: 
 a transcription promoter;    a DNA segment encoding a polypeptide according to  claim 1;  and    a transcription terminator;    wherein said cell expresses said polypeptide encoded by said DNA segment.    
     
     
         27 . A method of producing a polypeptide comprising: 
 culturing a cell into which has been introduced an expression vector comprising the following operably linked elements:    a transcription promoter;    a DNA segment encoding a polypeptide according to  claim 1;  and    a transcription terminator;    whereby said cell expresses said polypeptide encoded by said DNA segment; and    recovering said expressed polypeptide.    
     
     
         28 . An oligonucleotide probe or primer comprising at least 14 contiguous nucleotides of a polynucleotide of SEQ ID NO:10 or a sequence complementary to SEQ ID NO:10.  
     
     
         29 . A method for modulating free fatty acid metabolism by administering a pharmaceutically effective dose of a polypeptide according to  claim 1 .  
     
     
         30 . An isolated polynucleotide encoding a polypeptide selected from the group consisting of: 
 a) a polypeptide comprising a sequence of amino acid residues from amino acid residue 30 to amino acid residue 95 of SEQ ID NO:2;    b) a polypeptide comprising a sequence of amino acid residues from amino acid residue 30 to amino acid residue 96 of SEQ ID NO:2;    c) a polypeptide comprising a sequence of amino acid residues from amino acid residue 30 to amino acid residue 97 of SEQ ID NO:2;    d) a polypeptide comprising a sequence of amino acid residues from amino acid residue 98 to amino acid residue 243 of SEQ ID NO:2;    e) a polypeptide comprising a sequence of amino acid residues from amino acid residue 99 to amino acid residue 243 of SEQ ID NO:2; and    f) a polypeptide comprising a sequence of amino acid residues from amino acid residue 30 to amino acid residue 243 of SEQ ID NO:2.    
     
     
         31 . An isolated polynucleotide according to  claim 30 , wherein said polynucleotide is DNA.  
     
     
         32 . An expression vector comprising the following operably linked elements: 
 a transcription promoter;    a DNA segment encoding a polypeptide according to  claim 30;  and    a transcription terminator.    
     
     
         33 . An expression vector according to  claim 32 , wherein said DNA segment encodes a polypeptide covalently linked amino terminally or carboxy terminally to an affinity tag.  
     
     
         34 . An expression vector according to  claim 32  wherein said DNA segment further encodes a secretory signal sequence operably linked to said polypeptide.  
     
     
         35 . An expression vector according the  claim 32 , wherein said secretory signal sequence comprises residues 1-15 or 1-18 of SEQ ID NO:2.  
     
     
         36 . A cultured cell into which has been introduced an expression vector comprising the following operably linked elements: 
 a transcription promoter;    a DNA segment encoding a polypeptide according to  claim 30;  and    a transcription terminator;    wherein said cell expresses said polypeptide encoded by said DNA segment.    
     
     
         37 . A method of producing a polypeptide comprising: 
 culturing a cell into which has been introduced an expression vector comprising the following operably linked elements:    a transcription promoter;    a DNA segment encoding a polypeptide according to  claim 30;  and    a transcription terminator;    whereby said cell expresses said polypeptide encoded by said DNA segment; and    recovering said expressed polypeptide.