22196, a novel human aminopeptidase

Abstract

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

That which is claimed:  
     
         1 . A method for screening test compounds to identify a compound that binds to a polypeptide comprising an amino acid sequence selected from the group consisting of: 
 a) the amino acid sequence set forth in SEQ ID NO:1;    b) the amino acid sequence encoded by the cDNA insert of the plasmid deposited with the ATCC as Patent Deposit No. PTA-1662;    c) the amino acid sequence of a fragment of the amino acid sequence set forth in SEQ ID NO:1, wherein said fragment has endopeptidase activity and consists of at least 100 contiguous amino acids of the amino acid sequence set forth in SEQ ID NO:1; and    d) the amino acid sequence of a fragment of the amino acid sequence encoded by the cDNA insert of the plasmid deposited with the ATCC as Patent Deposit No. PTA-1662, wherein said fragment has endopeptidase activity and consists of at least 100 contiguous amino acids of the amino acid sequence encoded by the cDNA insert of the plasmid deposited with the ATCC as Patent Deposit No. PTA-1662;    said method comprising contacting the test compound with the polypeptide or a cell expressing the polypeptide under conditions suitable for binding of the compound to the polypeptide and determining whether a complex is formed between the test compound and the polypeptide.    
     
     
         2 . The method of  claim 1 , wherein said polypeptide comprises the amino acid sequence set forth in SEQ ID NO:1.  
     
     
         3 . A method for screening test compounds to identify a compound that binds to a polypeptide comprising the amino-acid sequence of a sequence variant of the amino acid sequence set forth in SEQ ID NO:1, wherein said sequence variant has endopeptidase activity and has at least 80% sequence identity with the amino acid sequence set forth in SEQ ID NO:1; said method comprising the steps of: 
 a) contacting the test compound with a cell expressing the polypeptide under conditions suitable for binding of the compound to the polypeptide; and    b) determining whether a complex is formed between the test compound and the polypeptide,    wherein said cell is selected from the group consisting of a lung cell, a colon cell, an osteoblast, and a dorsal root ganglion cell.    
     
     
         4 . The method of  claim 3 , wherein said polypeptide comprises the amino acid sequence of a sequence variant of the amino acid sequence set forth in SEQ ID NO:1, wherein said sequence variant has endopeptidase activity and has at least 90% sequence identity with the amino acid sequence set forth in SEQ ID NO:1  
     
     
         5 . The method of  claim 1 , wherein a cell expressing the polypeptide is contacted.  
     
     
         6 . The method of  claim 5 , wherein said cell is a dorsal root ganglion cell  
     
     
         7 . A method for screening test compounds to identify a compound that modulates the activity of a polypeptide comprising an amino acid sequence selected from the group consisting of: 
 a) the amino acid sequence set forth in SEQ ID NO:1;    b) the amino acid sequence encoded by the cDNA insert of the plasmid deposited with the ATCC as Patent Deposit No. PTA-1662;    c) the amino acid sequence of a fragment of the amino acid sequence set forth in SEQ ID NO:1, wherein said fragment has endopeptidase activity and consists of at least 100 contiguous amino acids of the amino acid sequence set forth in SEQ ID NO:1; and    d) the amino acid sequence of a fragment of the amino acid sequence encoded by the cDNA insert of the plasmid deposited with the ATCC as Patent Deposit No. PTA-1662, wherein said fragment has endopeptidase activity and consists of at least 100 contiguous amino acids of the amino acid sequence encoded by the cDNA insert of the plasmid deposited with the ATCC as Patent Deposit No. PTA-1662;    said method comprising contacting the test compound with the polypeptide or a cell expressing the polypeptide under conditions suitable for detecting the activity of the polypeptide and assaying the ability of the test compound to modulate the activity of the polypeptide.    
     
     
         8 . The method of  claim 7 , wherein said polypeptide comprises the amino acid sequence set forth in SEQ ID NO:1.  
     
     
         9 . The method of  claim 7 , wherein a cell expressing the polypeptide is contacted.  
     
     
         10 . The method of  claim 9 , wherein said cell is a dorsal root ganglion cell  
     
     
         11 . A method for screening test compounds to identify a compound that modulates the endopeptidase activity of a polypeptide comprising the amino acid sequence of a sequence variant of the amino acid sequence set forth in SEQ ID NO:1, wherein said sequence variant has endopeptidase activity and has at least 80% sequence identity with the amino acid sequence set forth in SEQ ID NO:1; said method comprising the steps of: 
 a) contacting the test compound with a cell expressing the polypeptide under conditions suitable for detecting the endopeptidase activity of the polypeptide; and    b) assaying the ability of the test compound to modulate the endopeptidase activity of the polypeptide    wherein said cell is selected from the group consisting of a lung cell, a colon cell, an osteoblast, and a dorsal root ganglion cell.    
     
     
         12 . The method of  claim 11 , wherein said polypeptide comprises the amino acid sequence of a sequence variant of the amino acid sequence set forth in SEQ ID NO:1, wherein said sequence variant has endopeptidase activity and has at least 90% sequence identity with the amino acid sequence set forth in SEQ ID NO:1  
     
     
         13 . A method for screening test compounds to identify a compound that binds to a nucleic acid molecule comprising a nucleotide sequence selected from the group consisting of: 
 a) the nucleotide sequence set forth in SEQ ID NO:2;    b) a nucleotide sequence encoding the amino acid sequence set forth in SEQ ID NO:1;    c) a nucleotide sequence having at least 80% sequence identity with the nucleotide sequence set forth in SEQ ID NO:2 wherein said nucleotide sequence having at least 80% sequence identity with the nucleotide sequence set forth in SEQ ID NO:2 encodes a polypeptide having endopeptidase activity; and    d) a nucleotide sequence having at least 80% sequence identity with the nucleotide sequence set forth in SEQ ID NO:2 wherein said nucleotide sequence having at least 80% sequence identity with the nucleotide sequence set forth in SEQ ID NO:2 encodes a polypeptide having endopeptidase activity;    said method comprising the steps of contacting the test compound with the nucleic acid molecule or a cell comprising the nucleic acid molecule under conditions suitable for the test compound to bind to the nucleic acid molecule and determining whether the test compound forms a complex with the nucleic acid molecule.    
     
     
         14 . A method for screening test compounds to identify a compound that modulates the level of a nucleic acid molecule comprising a nucleotide sequence selected from the group consisting of: 
 a) the nucleotide sequence set forth in SEQ ID NO:2;    b) a nucleotide sequence encoding the amino acid sequence set forth in SEQ ID NO:1;    c) a nucleotide sequence having at least 80% sequence identity with the nucleotide sequence set forth in SEQ ID NO:2 wherein said nucleotide sequence having at least 80% sequence identity with the nucleotide sequence set forth in SEQ ID NO:2 encodes a polypeptide having endopeptidase activity; and    d) a nucleotide sequence having at least 80% sequence identity with the nucleotide sequence set forth in SEQ ID NO:2 wherein said nucleotide sequence having at least 80% sequence identity with the nucleotide sequence set forth in SEQ ID NO:2 encodes a polypeptide having endopeptidase activity;    in a cell, said method comprising contacting the test compound with a cell containing the nucleic acid molecule under conditions suitable for the test compound to modulate the level of the nucleic acid molecule and then determining the level of the nucleic acid molecule.    
     
     
         15 . A method for modulating the activity of a polypeptide comprising an amino acid sequence selected from the group consisting of: 
 a) the amino acid sequence set forth in SEQ ID NO:1;    b) the amino acid sequence encoded by the cDNA insert of the plasmid deposited with the ATCC as Patent Deposit No. PTA-1662;    c) the amino acid sequence of a fragment of the amino acid sequence set forth in SEQ ID NO:1, wherein said fragment has endopeptidase activity and consists of at least 100 contiguous amino acids of the amino acid sequence set forth in SEQ ID NO:1; and    d) the amino acid sequence of a fragment of the amino acid sequence encoded by the cDNA insert of the plasmid deposited with the ATCC as Patent Deposit No. PTA-1662, wherein said fragment has endopeptidase activity and consists of at least 100 contiguous amino acids of the amino acid sequence encoded by the cDNA insert of the plasmid deposited with the ATCC as Patent Deposit No. PTA-1662;    said method comprising contacting the polypeptide or a cell expressing the polypeptide with a compound that modulates the activity of the polypeptide.    
     
     
         16 . The method of  claim 15 , wherein said polypeptide comprises the amino acid sequence set forth in SEQ ID NO:1.  
     
     
         17 . The method of  claim 15 , wherein a cell expressing the polypeptide is contacted.  
     
     
         18 . The method of  claim 17 , wherein said cell is a dorsal root ganglion cell  
     
     
         19 . A method for modulating the endopeptidase activity of a sequence variant of the amino acid sequence set forth in SEQ ID NO:1, wherein said sequence variant has endopeptidase activity and has at least 80% sequence identity with the amino acid sequence set forth in SEQ ID NO:1; said method comprising the steps of: 
 a) contacting the test compound with a cell expressing the polypeptide under conditions suitable for detecting the endopeptidase activity of the polypeptide; and    b) assaying the ability of the test compound to modulate the endopeptidase activity of the polypeptide    wherein said cell is selected from the group consisting of a lung cell, a colon cell, an osteoblast, and a dorsal root ganglion cell.    
     
     
         20 . The method of  claim 19 , wherein said polypeptide comprises the amino acid sequence of a sequence variant of the amino acid sequence set forth in SEQ ID NO:1, wherein said sequence variant has endopeptidase activity and has at least 90% sequence identity with the amino acid sequence set forth in SEQ ID NO:1