22012, a novel human carboxypeptidase

Abstract

The present invention relates to a newly identified human carboxypeptidase. The invention also relates to polynucleotides encoding the carboxypeptidase. The invention further relates to methods using the carboxypeptidase polypeptides and polynucleotides as a target for diagnosis and treatment in carboxypeptidase-related disorders. The invention further relates to drug-screening methods using the carboxypeptidase polypeptides and polynucleotides to identify agonists and antagonists for diagnosis and treatment. The invention further encompasses agonists and antagonists based on the carboxypeptidase polypeptides and polynucleotides. The invention further relates to procedures for producing the carboxypeptidase polypeptides and polynucleotides.

That which is claimed:  
     
         1 . An isolated nucleic acid molecule selected from the group consisting of: 
 a) a nucleic acid molecule comprising a nucleotide sequence having at least 85% sequence identity to the nucleotide sequence of SEQ ID NO:2, or the nucleotide sequence of the cDNA insert of the plasmid deposited with ATCC as Accession Number PTA-1643, wherein said nucleotide sequence encodes a polypeptide having carboxypeptidase activity;    b) a nucleic acid molecule comprising a fragment of at least 50 contiguous nucleotides of the nucleotide sequence of SEQ ID NO:2, or the nucleotide sequence of the cDNA insert of the plasmid deposited with ATCC as Accession Number PTA-1643,    c) a nucleic acid molecule which encodes a fragment of a polypeptide comprising the amino acid sequence of SEQ ID NO:1, or the amino acid sequence encoded by the cDNA insert of the plasmid deposited with the ATCC as Accession Number PTA-1643, wherein the fragment comprises at least 15 contiguous amino acids of SEQ ID NO:2, or the amino acid sequence encoded by the cDNA insert of the plasmid deposited with the ATCC as Accession Number PTA-1643,    d) a nucleic acid molecule which encodes a polypeptide having carboxypeptidase activity comprising the amino acid sequence of SEQ ID NO:1, or the amino acid sequence encoded by the cDNA insert of the plasmid deposited with the ATCC as Accession Number PTA-1643, wherein the nucleic acid molecule hybridizes to a nucleic acid molecule comprising the complement of SEQ ID NO:2 under stringent conditions; and    e) a nucleic acid molecule comprising the complement of a), b), c), or d).    
     
     
         2 . The isolated nucleic acid molecule of  claim 1 , comprising a nucleotide sequence having at least 95% sequence identity to the nucleotide sequence of SEQ ID NO:2, or the nucleotide sequence of the cDNA insert of the plasmid deposited with ATCC as Accession Number PTA-1643, wherein said nucleotide sequence encodes a polypeptide having carboxypeptidase activity, or a complement thereof.  
     
     
         3 . The nucleic acid molecule of  claim 1  further comprising vector nucleic acid sequences.  
     
     
         4 . The nucleic acid molecule of  claim 1  further comprising nucleic acid sequences encoding a heterologous polypeptide.  
     
     
         5 . A host cell which contains the nucleic acid molecule of  claim 3 .  
     
     
         6 . The host cell of  claim 5  which is a mammalian host cell.  
     
     
         7 . A non-human mammalian host cell containing the nucleic acid molecule of  claim 1 .  
     
     
         8 . An isolated polypeptide selected from the group consisting of: 
 a) a polypeptide having carboxypeptidase activity which is encoded by a nucleic acid molecule comprising a nucleotide sequence having at least 85% identity to a nucleic acid comprising the nucleotide sequence of SEQ ID NO:2, or the nucleotide sequence of the cDNA insert of the plasmid deposited with ATCC as Accession Number PTA-1643;    b) a polypeptide having carboxypeptidase activity, wherein the polypeptide is encoded by a nucleic acid molecule which hybridizes to a nucleic acid molecule comprising the complement of SEQ ID NO:2, or the nucleotide sequence of the cDNA insert of the plasmid deposited with ATCC as Accession Number PTA-1643 under stringent conditions;    c) a fragment of a polypeptide comprising the amino acid sequence of SEQ ID NO: 1, or the amino acid sequence encoded by the cDNA insert of the plasmid deposited with the ATCC as Accession Number PTA-1643, wherein the fragment comprises at least 15 contiguous amino acids of SEQ ID NO: 1; and    d) a polypeptide having at least 85% sequence identity to the amino acid sequence SEQ ID NO: 1, wherein the polypeptide has carboxypeptidase activity.    
     
     
         9 . The isolated polypeptide of  claim 8  comprising the amino acid sequence of SEQ ID NO:1.  
     
     
         10 . The polypeptide of  claim 8  further comprising heterologous amino acid sequences.  
     
     
         11 . An antibody which selectively binds to a polypeptide of  claim 8 .  
     
     
         12 . A method for producing a polypeptide selected from the group consisting a) a polypeptide comprising a fragment of the amino acid sequence of SEQ ID NO: 1, or the amino acid sequence encoded by the cDNA insert of the plasmid deposited with the ATCC as Accession Number PTA-1643, wherein the fragment comprises at least 15 contiguous amino acids of SEQ ID NO:1, or the amino acid sequence encoded by the cDNA insert of the plasmid deposited with the ATCC as Accession Number PTA-1643; 
 c) a polypeptide having carboxypeptidase activity, wherein the polypeptide is encoded by a nucleic acid molecule which hybridizes to a nucleic acid molecule comprising the complement of SEQ ID NO:2, or the nucleotide sequence of the cDNA insert of the plasmid deposited with ATCC as Accession Number PTA-1643,    d) a polypeptide having at least 85% sequence identity to the amino acid sequence of SEQ ID NO:2, wherein said polypeptide has carboxypeptidase activity; comprising culturing the host cell of  claim 5  under conditions in which the nucleic acid molecule is expressed.    
     
     
         13 . A method for detecting the presence of a polypeptide of  claim 8  in a sample, comprising: 
 a) contacting the sample with a compound which selectively binds to a polypeptide of claim  8 ; and  
 b) determining whether the compound binds to the polypeptide in the sample.  
 
     
     
         14 . The method of  claim 13 , wherein the compound which binds to the polypeptide is an antibody.  
     
     
         15 . A kit comprising a compound which selectively binds to a polypeptide of  claim 8  and instructions for use.  
     
     
         16 . A method for detecting the presence of a nucleic acid molecule of  claim 1  in a sample, comprising the steps of: 
 a) contacting the sample with a nucleic acid probe or primer which selectively hybridizes to the nucleic acid molecule; and  
 b) determining whether the nucleic acid probe or primer binds to a nucleic acid molecule in the sample.  
 
     
     
         17 . The method of  claim 16 , wherein the sample comprises mRNA molecules and is contacted with a nucleic acid probe.  
     
     
         18 . A kit comprising a compound which selectively hybridizes to a nucleic acid molecule of  claim 1  and instructions for use.  
     
     
         19 . A method for identifying a compound which binds to a polypeptide of  claim 8  comprising the steps of: 
 a) contacting a polypeptide, or a cell expressing a polypeptide of  claim 8  with a test compound; and  
 b) determining whether the polypeptide binds to the test compound.  
 
     
     
         20 . The method of  claim 19 , wherein the binding of the test compound to the polypeptide is detected by a method selected from the group consisting of: 
 a) detection of binding by direct detecting of test compound/polypeptide binding;    b) detection of binding using a competition binding assay;    c) detection of binding using an assay for carboxypeptidase activity.    
     
     
         21 . A method for modulating the activity of a polypeptide of  claim 8  comprising contacting a polypeptide or a cell expressing a polypeptide of  claim 8  with a compound which binds to the polypeptide in a sufficient concentration to modulate the activity of the polypeptide.  
     
     
         22 . A method for identifying a compound which modulates the activity of a polypeptide of  claim 8 , comprising: 
 a) contacting a polypeptide of  claim 8  with a test compound; and    b) determining the effect of the test compound on the activity of the polypeptide to thereby identify a compound that modulates the activity of the polypeptide.