US2004115700A1PendingUtilityA1

Human sel-10 polypeptides and polynucleotides that encode them

58
Priority: Dec 19, 1997Filed: Sep 2, 2003Published: Jun 17, 2004
Est. expiryDec 19, 2017(expired)· nominal 20-yr term from priority
Inventors:Mark E. GurneyAdele M. PauleyJinhe Li
C07K 14/4702C07K 2319/00C12N 15/11
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Claims

Abstract

The present invention provides isolated nucleic acid molecules comprising a polynucleotide encoding either of two alternative splice variants of human sel-10, one of which is expressed in hippocampal cells, and one of which is expressed in mammary cells. The invention also provides isolated sel-10 polypeptides and cell lines which express them in which Aβ processing is altered.

Claims

exact text as granted — not AI-modified
What is claimed is:  
     
         1 . An isolated nucleic acid molecule comprising a polynucleotide having a sequence at least 95% identical to a sequence selected from the group consisting of: 
 (a) a nucleotide sequence encoding a human sel-10 polypeptide having the complete amino acid sequence selected from the group consisting of SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:5, SEQ ID NO:6, and SEQ ID NO:7, or as encoded by the cDNA clone contained in ATCC Deposit No.98978;    (b) a nucleotide sequence encoding a human sel-10 polypeptide having the complete amino acid sequence selected from the group consisting of SEQ ID NO:8, SEQ ID NO:9, and SEQ ID NO:10, or as encoded by the cDNA clone contained in ATCC Deposit No. 98979; and    (c) a nucleotide sequence complementary to the nucleotide sequence of (a) or (b).    
     
     
         2 . An isolated nucleic acid molecule comprising polynucleotide which hybridizes under stringent conditions to a polynucleotide having the nucleotide sequence in (a), (b), or (c) of  claim 1 .  
     
     
         3 . The nucleic acid molecule of  claim 1 , wherein said polynucleotide of 1(a) encodes a human sel-10 polypeptide having the complete amino acid sequence of SEQ ID NO:3.  
     
     
         4 . The nucleic acid molecule of  claim 3 , wherein said polynucleotide molecule of 1(a) comprises the nucleotide sequence of residues 45-1928 of SEQ ID NO:1.  
     
     
         5 . The nucleic acid molecule of  claim 1 , wherein said polynucleotide of 1(a) encodes a human sel-10polypeptide having the complete amino acid sequence of SEQ ID NO:4.  
     
     
         6 . The nucleic acid molecule of  claim 5 , wherein said polynucleotide molecule of 1(a) comprises the nucleotide sequence of residues 150-1928 of SEQ ID NO:1.  
     
     
         7 . The nucleic acid molecule of  claim 1 , wherein said polynucleotide of 1(a) encodes a human sel-10 polypeptide having the complete amino acid sequence of SEQ ID NO:5.  
     
     
         8 . The nucleic acid molecule of  claim 7 , wherein said polynucleotide molecule of 1(a) comprises the nucleotide sequence of residues 267-1928 of SEQ ID NO:1.  
     
     
         9 . The nucleic acid molecule of  claim 1 , wherein said polynucleotide of 1(a) encodes a human sel-10 polypeptide having the complete amino acid sequence of SEQ ID NO:6.  
     
     
         10 . The nucleic acid molecule of  claim 9 , wherein said polynucleotide molecule of 1(a) comprises the nucleotide sequence of residues 291-1928 of SEQ ID NO:1.  
     
     
         11 . The nucleic acid molecule of  claim 1 , wherein said polynucleotide of 1(a) encodes a human sel-10 polypeptide having the complete amino acid sequence of SEQ ID NO:7.  
     
     
         12 . The nucleic acid molecule of  claim 11 , wherein said polynucleotide molecule of 1(a) comprises the nucleotide sequence of residues 306-1928 of SEQ ID NO:1.  
     
     
         13 . The nucleic acid molecule of  claim 1 , wherein said polynucleotide of 1(b) encodes a human sel-10 polypeptide having the complete amino acid sequence of SEQ ID NO:8.  
     
     
         14 . The nucleic acid molecule of  claim 13  wherein said polynucleotide molecule of 1(b) comprises the nucleotide sequence of residues 180-1949 of SEQ ID NO:2.  
     
     
         15 . The nucleic acid molecule of  claim 1 , wherein said polynucleotide of 1(b) encodes a human sel-10polypeptide having the complete amino acid sequence of SEQ ID NO:9.  
     
     
         16 . The nucleic acid molecule of  claim 15 , wherein said polynucleotide molecule of 1(b) comprises the nucleotide sequence of residues 270-1949 of SEQ ID NO:2.  
     
     
         17 . The nucleic acid molecule of  claim 1 , wherein said polynucleotide of 1(b) encodes a human sel-10 polypeptide having the complete amino acid sequence of SEQ ID NO:10.  
     
     
         18 . The nucleic acid molecule of  claim 17 , wherein said polynucleotide molecule of 1(b) comprises the nucleotide sequence of residues 327-1949 of SEQ ID NO:2.  
     
     
         19 . A vector comprising the nucleic acid molecule of  claim 1 .  
     
     
         20 . The vector of  claim 19 , wherein said nucleic acid molecule of  claim 1  is operably linked to a promoter for the expression of a sel-10 polypeptide.  
     
     
         21 . A host cell comprising the vector of  claim 19 .  
     
     
         22 . The host cell of  claim 21 , wherein said host is a eukaryotic host.  
     
     
         23 . A method of obtaining a sel-10 polypeptide comprising culturing the host cell of  claim 22  and isolating said sel-10 polypeptide.  
     
     
         24 . An isolated sel-10 polypeptide comprising 
 (a) an amino acid sequence selected from the group consisting of SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:5, SEQ ID NO:6, and SEQ ID NO:7, or as encoded by the cDNA clone contained in ATCC Deposit No. 98978;    (b) an amino acid sequence selected from the group consisting of SEQ ID NO:8, SEQ ID NO:9, and SEQ ID NO:10, or as encoded by the cDNA clone contained in ATCC Deposit No. 98979.    
     
     
         25 . The isolated sel-10 polypeptide of  claim 24 , wherein said polypeptide comprises the amino acid sequence of SEQ ID NO:3.  
     
     
         26 . The isolated sel-10 polypeptide of  claim 24 , wherein said polypeptide comprises the amino acid sequence of SEQ ID NO:4.  
     
     
         27 . The isolated sel-10 polypeptide of  claim 24 , wherein said polypeptide comprises the amino acid sequence of SEQ ID NO:5.  
     
     
         28 . The isolated sel-10 polypeptide of  claim 24 , wherein said polypeptide comprises the amino acid sequence of SEQ ID NO:6.  
     
     
         29 . The isolated sel-10 polypeptide of  claim 24 , wherein said polypeptide comprises the amino acid sequence of SEQ ID NO:7.  
     
     
         30 . The isolated sel-10 polypeptide of  claim 24 , wherein said polypeptide comprises the amino acid sequence of SEQ ID NO:8.  
     
     
         31 . The isolated sel-10 polypeptide of  claim 24 , wherein said polypeptide comprises the amino acid sequence of SEQ ID NO:9.  
     
     
         32 . The isolated sel-10 polypeptide of  claim 24 , wherein said polypeptide comprises the amino acid sequence of SEQ ID NO:10.  
     
     
         33 . An isolated antibody that binds specifically to the sel-10 polypeptide of  claim 24 .  
     
     
         34 . A cell line having altered Aβ processing that expresses any of the sel-10 isolated nucleic acid molecules of  claim 1 .  
     
     
         35 . The cell line of  claim 34 , wherein said Aβ processing is increased.  
     
     
         36 . The cell line of  claim 34 , wherein said Aβ processing is decreased.  
     
     
         37 . The cell line of  claim 34 , wherein said cell line is 6myc-N-sel10/2.  
     
     
         38 . The cell line of  claim 34 , wherein said cell line is 6myc-N-sel10/6.  
     
     
         39 . A method for the identification of an agent capable of altering the ratio of Aβ 1-40 /Aβ 1-40 +Aβ 1-42  produced in any of the cell lines of claims  34 ,  37 , and  38 , comprising the steps of: 
 (a) obtaining a test culture and a control culture of said cell line;  
 (b) contacting said test culture with a test agent;  
 (c) measuring the levels of Aβ 1-40  and Aβ 1-42  produced by said test culture of step (b) and said control culture;  
 (d) calculating the ratio of Aβ 1-40 /Aβ 1-40 +Aβ 1-42  for said test culture and said control culture from the levels of Aβ 1-40  and Aβ 1-42  measured in step (c); and  
 (e) comparing the ratio of Aβ 1-40 /Aβ 1-40 +Aβ 1-42  measured for said test culture and said control culture in step (d);  
 whereby a determination that the ratio of Aβ 1-40 /Aβ 1-40 +Aβ 1-42  for said test culture is higher or lower than ratio of Aβ 1-40 /Aβ 1-40 +Aβ 1-42  for said control culture indicates that said test agent has altered the ratio of Aβ 1-40 /Aβ 1-40 +Aβ 1-42 .  
 
     
     
         40 . The method of  claim 39 , wherein said ratio of Aβ 1-40 /Aβ 1-40 +Aβ 1-42  is increased by said test agent.  
     
     
         41 . The method of  claim 39 , wherein said ratio of Aβ 1-40 /Aβ 1-40 +Aβ 1-42  is decreased by said test agent.

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