Adipocyte complement related protein homolog zacrp3

Abstract

The present invention relates to polynucleotide and polypeptide molecules for zacrp3, a novel member of the family of proteins bearing a collagen-like domain and a Clq 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 zacrp3 polypeptides.

What is claimed is:  
     
         1 . An isolated polypeptide comprising a sequence of amino acid residues that is at least 75% identical in amino acid sequence to residues 51-246 of SEQ ID NO:2, wherein said sequence comprises: 
 Gly-Xaa-Xaa or Gly-Xaa-Pro repeats forming a collagen domain, wherein Xaa is any amino acid; and    a carboxyl-terminal Clq domain comprising 10 beta strands.    
     
     
         2 . An isolated polypeptide according to  claim 1 , wherein said polypeptide that is at least 90% identical in amino acid sequence to residues 23-246 of SEQ ID NO:2.  
     
     
         3 . An isolated polypeptide according to  claim 1 , wherein said collagen domain consists of 15 Gly-Xaa-Xaa repeats and 6 Gly-Xaa-Pro repeats.  
     
     
         4 . An isolated polypeptide according to  claim 1 , wherein said carboxyl-terminal Clq domain comprises the sequence of SEQ ID NO:10.  
     
     
         5 . An isolated polypeptide according to  claim 1 , wherein said carboxy-terminal Clq domain comprises amino acid residues 119-123, 140-142, 148-151, 155-158, 161-173, 175-182, 190-197, 200-212, 217-222 and 236-241 of SEQ ID NO:2.  
     
     
         6 . An isolated polypeptide according to  claim 1 , wherein any differences between said polypeptide and SEQ ID NO:2 are due to conservative amino acid substitutions.  
     
     
         7 . An isolated polypeptide according to  claim 1 , wherein said polypeptide specifically binds with an antibody that specifically binds with a polypeptide consisting of the amino acid sequence of SEQ ID NO:2.  
     
     
         8 . An isolated polypeptide according to  claim 2 , wherein said polypeptide comprises residues 23-246 of SEQ ID NO:2.  
     
     
         9 . An isolated polypeptide according to  claim 1 , wherein said collagen domain consists of amino acid residues 51-113 of SEQ ID NO:2.  
     
     
         10 . An isolated polypeptide according to  claim 1 , wherein said carboxy-terminal Clq domain consists of amino acid residues 114-246 of SEQ ID NO:2.  
     
     
         11 . An isolated polypeptide according to  claim 1 , covalently linked at the amino or carboxyl terminus to a moiety selected from the group consisting of affinity tags, toxins, radionucleotides, enzymes and fluorophores.  
     
     
         12 . An isolated polypeptide selected from the group consisting of: 
 a) a polypeptide consisting of a sequence of amino acid residues that is 80% identical in amino acid sequence to amino acid residue 51 to amino acid residue 113 of SEQ ID NO:2, said polypeptide consisting of Gly-Xaa-Xaa and Gly-Xaa-Pro repeats forming a collagen domain;    b) a polypeptide consisting of a sequence of amino acid residues that is 80% identical in amino acid sequence to amino acid residue 114 to amino acid residue 246 of SEQ ID NO:2 comprising the sequence of SEQ ID NO:5; and    c) a polypeptide consisting of a sequence of amino acid residues that is 80% identical in amino acid sequence to amino acid residue 51 to 246 of SEQ ID NO:2, said polypeptide consisting of Gly-Xaa-Xaa and Gly-Xaa-Pro repeats forming a collagen domain and comprising the sequence of SEQ ID NO:5.    
     
     
         13 . A fusion protein comprises a first portion and a second portion joined by a peptide bond, said first portion consisting of a polypeptide selected from the group consisting of: 
 a) a polypeptide comprising a sequence of amino acid residues that is at least 75% identical in amino acid sequence to amino acid residue 51 to amino acid residue 246 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 1 to amino acid residue 246;    c a portion of the zacrp3 polypeptide of SEQ ID NO:2, comprising the collagen-like domain or a portion of the collagen-like domain capable of dimerization or oligomerization;    d) a portion of the zacrp3 polypeptide of SEQ ID NO:2, comprising the Clq domain or an active portion of the Clq domain; or    e) a portion of the zacrp3 polypeptide of SEQ ID NO:2 comprising of the collagen-like domain and the Clq domain; and    said second portion comprising another polypeptide.    
     
     
         14 . A fusion protein according to  claim 13 , wherein said first portion is selected from the group consisting of: 
 a) a polypeptide consisting of the sequence of amino acid residue 51 to amino acid residue 113 of SEQ ID NO:2;    b) a polypeptide consisting of the sequence of amino acid residue 114 to amino acid residue 246 of SEQ ID NO:2;    c) a polypeptide consisting of the sequence of amino acid residue 51 to 246 of SEQ ID NO:2.    
     
     
         15 . A fusion protein according to  claim 13 , wherein said second portion comprises a collagen domain or a Clq domain from an adipocyte complement related protein.  
     
     
         16 . A polypeptide according to  claim 1;  in combination with a pharmaceutically acceptable vehicle.  
     
     
         17 . An antibody or antibody fragment that specifically binds to a polypeptide according to  claim 1 .  
     
     
         18 . An antibody according to  claim 17 , wherein said antibody is selected from the group consisting of: 
 a) polyclonal antibody;    b) murine monoclonal antibody;    c) humanized antibody derived from b); and    d) human monoclonal antibody.    
     
     
         19 . An antibody fragment according to  claim 17 , wherein said antibody fragment is selected from the group consisting of F(ab′), F(ab), Fab′, Fab, Fv, scFv, and minimal recognition unit.  
     
     
         20 . An anti-idiotype antibody that specifically binds to said antibody of  claim 17 .  
     
     
         21 . An isolated polynucleotide encoding a polypeptide comprising a sequence of amino acid residues that is at least 75% identical in amino acid sequence to residues 51-246 of SEQ ID NO:2, wherein said sequence comprises: 
 Gly-Xaa-Xaa or Gly-Xaa-Pro repeats forming a collagen domain, wherein Xaa is any amino acid; and    a carboxyl-terminal Clq domain comprising 10 beta strands.    
     
     
         22 . An isolated polynucleotide according to  claim 21 , wherein said polypeptide is at least 90% identical in amino acid sequence to residues 23-246 of SEQ ID NO:2.  
     
     
         23 . An isolated polynucleotide according to  claim 21 , wherein said collagen domain consists of 15 Gly-Xaa-Xaa repeats and 6 Gly-Xaa-Pro repeats.  
     
     
         24 . An isolated polynucleotide according to  claim 21 , wherein said carboxyl-terminal Clq domain comprises the sequence of SEQ ID NO:5.  
     
     
         25 . An isolated polynucleotide according to  claim 21 , wherein said carboxy-terminal Clq domain comprises amino acid residues 119-123, 140-142, 148-151, 155-158, 161-173, 175-182, 190-197, 200-212, 217-222 and 236-241 of SEQ ID NO:2.  
     
     
         26 . An isolated polynucleotide according to  claim 21 , wherein any differences between said polypeptide and SEQ ID NO:2 are due to conservative amino acid substitutions.  
     
     
         27 . An isolated polynucleotide according to  claim 21 , wherein said polypeptide specifically binds with an antibody that specifically binds with a polypeptide consisting of the amino acid sequence of SEQ ID NO:2.  
     
     
         28 . An isolated polynucleotide according to  claim 21 , wherein said collagen domain comprises amino acid residues 51-113 of SEQ ID NO:2.  
     
     
         29 . An isolated polynucleotide according to  claim 21 , wherein said carboxy-terminal Clq domain consists of amino acid residues 114-246 of SEQ ID NO:2.  
     
     
         30 . An isolated polynucleotide according to  claim 21 , wherein said polypeptide comprises residues 23-246 of SEQ ID NO:2.  
     
     
         31 . An isolated polynucleotide selected from the group consisting of, 
 a) a sequence of nucleotides from nucleotide 1 to nucleotide 1696 of SEQ ID NO:1;    b) a sequence of nucleotides from nucleotide 69 to nucleotide 806 of SEQ ID NO:1;    c) a sequence of nucleotides from nucleotide 135 to nucleotide 806 of SEQ ID NO:1;    d) a sequence of nucleotides from nucleotide 219 to nucleotide 806 of SEQ ID NO:1;    e) a sequence of nucleotides from nucleotide 408 to nucleotide 806 of SEQ ID NO:1;    f) a sequence of nucleotides from nucleotide 69 to nucleotide 407 of SEQ ID NO:1;    g) a sequence of nucleotides from nucleotide 135 to nucleotide 407 of SEQ ID NO:1;    h) a sequence of nucleotides from nucleotide 219 to nucleotide 407 of SEQ ID NO:1;    i) a polynucleotide encoding a polypeptide, said polypeptide consisting of a sequence of amino acid residues that is at least 75% identical to a polypeptide consisting of the amino acid sequence of residues 51 to 113 of SEQ ID NO:2;    j) a polynucleotide encoding a polypeptide, said polypeptide consisting of a sequence of amino acid residues that is at least 75% identical to a polypeptide consisting of the amino acid sequence of residues 114 to 246 of SEQ ID NO:2;    k) a polynucleotide encoding a polypeptide, said polypeptide consisting of a sequence of amino acid residues that is at least 75% identical to a polypeptide consisting of the amino acid sequence of residues 51 to 246 of SEQ ID NO:2;    l) a polynucleotide encoding a polypeptide consisting of a sequence of amino acid residues that is at least 75% identical to a polypeptide consisting of the amino acid sequence of residues 23 to 113 of SEQ ID NO:2;    m) a polynucleotide that remains hybridized following stringent wash conditions to a polynucleotide consisting of the nucleotide sequence of SEQ ID NO:1, or the complement of SEQ ID NO:1;    n) nucleotide sequences complementary to a), b), c), d), e), f), g), h), i), j), k), l) or m) and    o) degenerate nucleotide sequences of i), j), k) or l).    
     
     
         32 . An isolated polynucleotide encoding a fusion protein comprising 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 75% identical in amino acid sequence to amino acid residues 51 to 246 of SEQ ID NO:2;    b) a polypeptide comprising the sequence of amino acid residues 1 to 246 of SEQ ID NO:2;    c) a polypeptide comprising the sequence of amino acid residues 23 to 246 of SEQ ID NO:2;    d) a polypeptide comprising the sequence of amino acid residues 23 to 113 of SEQ ID NO:2;    e) a polypeptide comprising the sequence of amino acid residues 1 to 113 of SEQ ID NO:2;    f) a portion of a polypeptide of SEQ ID NO:2 comprising the collagen-like domain or a portion of the collagen-like domain capable of dimerization or oligomerization;    g) a portion of the polypeptide of SEQ ID NO:2 containing the Clq domain; or    h) a portion of the polypeptide of SEQ ID NO:2 including the collagen-like domain and the Clq domain; and    said second portion comprising another polypeptide.    
     
     
         33 . An isolated polynucleotide consisting of the sequence of nucleotide 1 to nucleotide 738 of SEQ ID NO:10.  
     
     
         34 . 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.    
     
     
         35 . An expression vector according to  claim 34 , wherein said DNA segment encodes a polypeptide that is at least 90% identical in amino acid sequence to residues 23-246 of SEQ ID NO:2.  
     
     
         36 . An expression vector according to  claim 34 , wherein said collagen domain consists of 15 Gly-Xaa-Xaa repeats and 6 Gly-Xaa-Pro repeats.  
     
     
         37 . An expression vector according to  claim 34 , wherein said carboxyl-terminal Clq domain comprises the sequence of SEQ ID NO:10.  
     
     
         38 . An expression vector according to  claim 34 , wherein said carboxy-terminal Clq domain comprises amino acid residues 119-123, 140-142, 148-151, 155-158, 161-173, 175-182, 190-197, 200-212, 217-222 and 236-241 of SEQ ID NO:2.  
     
     
         39 . An expression vector according to  claim 34 , wherein any differences between said polypeptide and SEQ ID NO:2 are due to conservative amino acid substitutions.  
     
     
         40 . An expression vector according to  claim 34 , wherein said polypeptide specifically binds with an antibody that specifically binds with a polypeptide consisting of the amino acid sequence of SEQ ID NO:2.  
     
     
         41 . An expression vector according to  claim 34 , wherein said collagen domain consists of amino acid residues 51-113 of SEQ ID NO:2.  
     
     
         42 . An expression vector according to  claim 34 , wherein said Clq domain consists of amino acid residues 114-246 of SEQ ID NO:2.  
     
     
         43 . An expression vector according to  claim 34 , wherein said DNA segment encodes a polypeptide comprising residues 23-246 of SEQ ID NO:2.  
     
     
         44 . An expression vector according to  claim 34 , wherein said DNA segment encodes a polypeptide covalently linked at the amino or carboxyl terminus to an affinity tag.  
     
     
         45 . An expression vector according to  claim 34  wherein said DNA segment further encodes a secretory signal sequence operably linked to said polypeptide.  
     
     
         46 . An expression vector according the  claim 34 , wherein said secretory signal sequence comprises residues 1-22 of SEQ ID NO:2.  
     
     
         47 . A cultured cell into which has been introduced an expression vector according to  claim 34 , wherein said cell expresses said polypeptide encoded by said DNA segment.  
     
     
         48 . A method of producing a polypeptide comprising: 
 culturing a cell into which has been introduced an expression vector according to  claim 34;     whereby said cell expresses said polypeptide encoded by said DNA segment; and    recovering said expressed polypeptide.