2786, 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 . An isolated polypeptide having an amino acid sequence selected from the group consisting of: 
 (a) the amino acid sequence shown in SEQ ID NO:1;    (b) an amino acid sequence encoded by the cDNA insert of the plasmid deposited with ATCC as Patent Deposit No. PTA-2811;    (c) an amino acid sequence of an allelic variant of the amino acid sequence shown in SEQ ID NO:1, wherein said allelic variant has aminopeptidase activity;    (d) an amino acid sequence of an allelic variant of the amino acid sequence encoded by the cDNA contained in ATCC Deposit No. PTA-2811, wherein said allelic variant has aminopeptidase activity;    (e) an amino acid sequence of a sequence variant of the amino acid sequence shown in SEQ ID NO:1, wherein the sequence variant is encoded by a nucleic acid molecule hybridizing to the nucleotide sequence shown in SEQ ID NO:2 under stringent conditions, and the sequence variant has aminopeptidase activity;    (f) an amino acid sequence of a sequence variant of the amino acid sequence encoded by the cDNA insert of the plasmid deposited with ATCC as Patent Deposit No. PTA-2811, wherein the sequence variant is encoded by a nucleic acid molecule hybridizing under stringent conditions to the cDNA insert of the plasmid deposited with ATCC as Patent Deposit No. PTA-2811, and the sequence variant has aminopeptidase activity;    (g) a fragment of the amino acid sequence shown in SEQ ID NO:1, wherein said fragment is encoded by a nucleotide sequence that is at least 500 nucleotides in length;    (h) a fragment of the amino acid sequence encoded by the cDNA insert of the plasmid deposited with ATCC as Patent Deposit No. PTA-2811, wherein said fragment is encoded by a nucleotide sequence that is at least 500 nucleotides in length;    (i) an amino acid sequence of the mature polypeptide from about amino acid 6 to the last amino acid shown in SEQ ID NO:1;    (j) an amino acid sequence of the mature polypeptide from about amino acid 6 to the last amino acid encoded by the cDNA insert of the plasmid deposited with ATCC as Patent Deposit No. PTA-2811;    (k) an amino acid sequence of a polypeptide having at least 60% sequence identity to the amino acid sequence of SEQ ID NO:1, wherein said polypeptide has aminopeptidase activity; and    (l) an amino acid sequence of an epitope bearing region of any one of the amino acid sequences of (a), (b), (c), (d), (e), (f), (g), (h), (i), (j), or (k).    
     
     
         2 . An isolated nucleic acid molecule having a nucleotide sequence selected from the group consisting of: 
 (a) a nucleotide sequence encoding an amino acid sequence of a sequence variant of the amino acid sequence shown in SEQ ID NO:1 that hybridizes to the nucleotide sequence shown in SEQ ID NO:2 under stringent conditions, wherein said sequence variant has aminopeptidase activity;    (b) a nucleotide sequence encoding the amino acid sequence of a sequence variant of the amino acid sequence encoded by the cDNA insert of the plasmid deposited with ATCC as Patent Deposit No. PTA-2811, wherein the nucleotide sequence hybridizes to the nucleotide sequence shown in SEQ ID NO:2 under stringent condition and the sequence variant has aminopeptidase activity;    (c) a nucleotide sequence that is at least 60% identical to the nucleotide sequence of SEQ ID NO:2 or the nucleotide sequence of the cDNA insert of the plasmid deposited with ATCC as Patent Deposit No. PTA-2811, wherein said nucleotide sequence encodes a polypeptide having aminopeptidase activity;    (d) a nucleotide sequence encoding a fragment of the amino acid sequence shown in SEQ ID NO:1 or the amino acid sequence encoded by the cDNA insert of the plasmid deposited with ATCC as Patent Deposit No. PTA-2811, wherein the fragment comprises at least 15 contiguous amino acids of the amino acid sequence shown in SEQ ID NO:1 or the amino acid sequence encoded by the cDNA insert of the plasmid deposited with ATCC as Patent Deposit No. PTA-2811; and    (e) a nucleotide sequence complementary to any of the nucleotide sequences in (a), (b), (c), (d).    
     
     
         3 . The nucleic acid molecule of  claim 2 , wherein said nucleotide sequence selected from the group consisting of: 
 (a) the nucleotide sequence shown in SEQ ID NO:2;    (b) the nucleotide sequence of the cDNA insert of the plasmid deposited with ATCC as Patent Deposit No: PTA-2811;    (c) a nucleotide sequence encoding the amino acid sequence shown in SEQ ID NO:1;    (d) a nucleotide sequence encoding the amino acid sequence encoded by the cDNA insert of the plasmid deposited with ATCC as Patent Deposit No. PTA-2811; and    (e) a nucleotide sequence complementary to any of the nucleotide sequences in (a), (b), (c), or (d).    
     
     
         4 . The nucleic acid molecule of  claim 1 , further comprising vector nucleic acid sequences.  
     
     
         5 . The nucleic acid molecule of  claim 1 , further comprising a nucleotide sequence encoding a heterologous polypeptide  
     
     
         6 . A host cell containing the nucleic acid molecule of  claim 1 .  
     
     
         7 . The host cell of  claim 6 , wherein said host cell is a mammalian host cell.  
     
     
         8 . A non-human mammalian host cell containing the nucleic acid molecule of  claim 1 .  
     
     
         9 . A method for producing a polypeptide selected from the group consisting of: 
 (a) the amino acid sequence shown in SEQ ID NO:1;    (b) an amino acid sequence encoded by the cDNA insert of the plasmid deposited with ATCC as Patent Deposit No. PTA-2811;    (c) an amino acid sequence of an allelic variant of the amino acid sequence shown in SEQ ID NO:1, wherein said allelic variant has aminopeptidase activity;    (d) an amino acid sequence of an allelic variant of the amino acid sequence encoded by the cDNA contained in ATCC Deposit No. PTA-2811, wherein said allelic variant has aminopeptidase activity;    (e) an amino acid sequence of a sequence variant of the amino acid sequence shown in SEQ ID NO:1, wherein the sequence variant has aminopeptidase activity and is encoded by a nucleic acid molecule hybridizing to the nucleotide sequence shown in SEQ ID NO:2 under stringent conditions;    (f) an amino acid sequence of a sequence variant of the amino acid sequence encoded by the cDNA insert of the plasmid deposited with ATCC as Patent Deposit No. PTA-2811, wherein the sequence variant has aminopeptidase activity and is encoded by a nucleic acid molecule hybridizing under stringent conditions to the cDNA insert of the plasmid deposited with ATCC as Patent Deposit No. PTA-2811;    (g) a fragment of the amino acid sequence shown in SEQ ID NO:1, wherein said fragment is encoded by a nucleotide sequence that is at least 500 nucleotides in length;    (h) a fragment of the amino acid sequence encoded by the cDNA insert of the plasmid deposited with ATCC as Patent Deposit No. PTA-2811, wherein said fragment is encoded by a nucleotide sequence that is at least 500 nucleotides in length;    (i) the amino acid sequence of the mature polypeptide from about amino acid 6 to the last amino acid shown in SEQ ID NO:1;    (j) the amino acid sequence of the mature polypeptide from about amino acid 6 to the last amino acid encoded by the cDNA insert of the plasmid deposited with ATCC as Patent Deposit No. PTA-2811;    (k) an amino acid sequence of a polypeptide having at least 60% sequence identity to the amino acid sequence of SEQ ID NO:1, wherein said polypeptide has aminopeptidase activity; and    (l) an amino acid sequence of an epitope bearing region of any one of the amino acid sequences of (a), (b), (c), (d), (e), (f), (g), (h), (i), (j), or (k) comprising culturing the host cell of  claim 6  under conditions in which the nucleic acid molecule is expressed.    
     
     
         10 . A method for detecting the presence of any of the polypeptides of  claim 1  in a sample, said method comprising: 
 (a) contacting said sample with an agent that specifically binds to a polypeptide of  claim 1;  and  
 (b) determining whether the compound binds to the polypeptide in the sample.  
 
     
     
         11 . The method of  claim 10 , wherein said agent is an antibody.  
     
     
         12 . A kit comprising reagents that selectively binds to a polypeptide of  claim 1  and instructions for use.  
     
     
         13 . A method for detecting the presence of a nucleic acid molecule of  claim 2  in a sample, comprising the steps of 
 (a) contacting said sample with an agent that selectively binds to the nucleic acid molecule in the sample; and  
 (b) determining whether the agent binds to a nucleic acid molecule in the sample.  
 
     
     
         14 . The method of  claim 13 , wherein the sample comprises mRNA molecules and the agent is a nucleic acid probe or primer.  
     
     
         15 . A kit having a compound which selectively hybridizes to a nucleic acid molecule of  claim 2  and instructions for use.  
     
     
         16 . A method for identifying a compound that binds to a polypeptide of  claim 1 , comprising the steps of: 
 (a) contacting a polypeptide of  claim 1 , or a cell expressing a polypeptide of  claim 1 , with a test compound; and    (b) determining whether the polypeptide binds to the test compound.    
     
     
         17 . The method of  claim 16 , 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 complex formation;    (b) detection of binding using a competition binding assay; and    (c) detection of binding using an assay for 2786-mediated protease activity.    
     
     
         18 . The method of  claim 16 , wherein the cell expressing the polypeptide is selected from the group consisting of: osteoclasts and breast tumor cells.  
     
     
         19 . A method for modulating the activity of a polypeptide of  claim 1  comprising contacting a polypeptide of  claim 1  or a cell expressing a polypeptide of  claim 1  with a compound that binds to the polypeptide under conditions such that the activity of the polypeptide is modulated.  
     
     
         20 . The method of  claim 19 , wherein the cell expressing the polypeptide is selected from the group consisting of: osteoblasts and breast tumor cells.  
     
     
         21 . A method for identifying a compound that modulates the activity of a polypeptide of  claim 1  comprising the steps of: 
 (a) contacting a polypeptide of  claim 1  or a cell expressing a polypeptide of  claim 1  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.  
 
     
     
         22 . The method of  claim 21 , wherein said compound is an antibody.  
     
     
         23 . The method of  claim 21 , wherein said polypeptide comprises heterologous amino acid sequences.  
     
     
         24 . The method of  claim 21 , wherein said compound inhibits binding of an agent to the polypeptide; said method further comprising combining the compound to be tested and the agent with the polypeptide under conditions suitable for binding of said agent thereto, and detecting the formation of a complex between said polypeptide and said agent, whereby inhibition of complex formation by said compound is indicative that said compound inhibits binding of said agent to said polypeptide.  
     
     
         25 . The method of  claim 24 , wherein said polypeptide can mediate cellular signaling or a cellular response, and the formation of a complex is monitored by detecting a signaling activity or cellular response of said polypeptide in response thereto.  
     
     
         26 . The method of  claim 21 , wherein said compound is an inhibitor of said polypeptide; said method further comprising assessing the ability of said compound to inhibit the activity of the polypeptide, wherein said activity is aminopeptidase activity and inhibition of said aminopeptidase activity by said agent is indicative that said agent is an inhibitor.  
     
     
         27 . The method of  claim 26 , wherein said polypeptide comprises heterologous amino acid sequences.  
     
     
         28 . The method of  claim 19 , wherein said modulation is in a cell selected from the group consisting of: osteoclasts and breast tumor cells.  
     
     
         29 . The method of  claim 19 , wherein said modulation is in a patient having a disorder involving brain, breast, fetal kidney, fetal liver, or bone mass.  
     
     
         30 . The method of  claim 19 , wherein said modulation is in a patient having abnormal or undesirable bone mass development.  
     
     
         31 . The method of  claim 19 , wherein said modulation is in a patient having breast cancer.