Viral proteins capable of binding LAR

Abstract

A poxvirus protein designated A41L binds to leukocyte common-antigen-related protein (LAR). A41L is a secreted protein that can be purified from the culture supernatant of cells infected with certain poxviruses, or produced using recombinant DNA techniques. A41L polypeptides and LAR polypeptides, and nucleic acids encoding them, are provided herein. Also provided are methods of using such polypeptides and nucleic acids.

1 . A purified oligomer comprising from two to four A41L polypeptides, wherein each of the A41L polypeptides comprises an amino acid sequence that is at least 80% identical to a sequence selected from the group consisting of: 
 a) a polypeptide comprising amino acids 25 to 208 of SEQ ID NO:2, inclusive;    b) a polypeptide comprising amino acids 26 to 209 of SEQ ID NO:3, inclusive;    c) a polypeptide comprising amino acids 26 to 209 of SEQ ID NO:4, inclusive;    d) a polypeptide comprising amino acids 26 to 209 of SEQ ID NO:5, inclusive;    e) a polypeptide comprising amino acids 26 to 209 of SEQ ID NO:6, inclusive;    f) a polypeptide comprising amino acids 26 to 208 of SEQ ID NO:7, inclusive;    g) a polypeptide comprising amino acids 26 to 208 of SEQ ID NO:8, inclusive; and    h) a polypeptide comprising amino acids 26 to 208 of SEQ ID NO:9, inclusive, wherein said A41L oligomer binds LAR.,    
     
     
         2 . A pharmaceutical composition comprising an oligomer of  claim 1  and a pharmaceutically acceptable diluent, excipient, or carrier.  
     
     
         3 . A method of treating inflammation, comprising administering an effective amount of an antagonist of LAR to a mammal afflicted with an inflammatory condition, wherein said antagonist of LAR is selected from the group consisting of: 
 a) a polypeptide comprising amino acids x to y, wherein x represents an integer from 1 to 20, inclusive, and y represents an integer from 960 to 965, inclusive, of SEQ ID NO:10;    b) a polypeptide comprising amino acids x to y, wherein x represents an integer from 1 to 20, inclusive, and y represents an integer from 302 to 312, inclusive, of SEQ ID NO:10;    c) a polypeptide comprising amino acids x to y, wherein x represents an integer from 1 to 20, inclusive, and y represents an integer from 296 to 306, inclusive, of SEQ ID NO:12;    d) fragments of the extracellular domain of LAR that are capable of binding A41L;    e) antagonistic antibodies to LAR; and    f) compounds that bind LAR and inhibit the binding of A41L thereto.    
     
     
         4 . The method of  claim 3 , wherein said mammal is a human.  
     
     
         5 . A method for identifying compounds that alter one or more biological activities of LAR, comprising: 
 (a) mixing a test compound with an A41L polypeptide selected from the group consisting of 
 (i) a polypeptide comprising amino acids x to y of SEQ ID NO:2, wherein x represents an integer from 1 to 25, inclusive, and y represents an integer from 208 to 218, inclusive, of SEQ ID NO:2;  
 (ii) a polypeptide comprising amino acids x to y of SEQ ID NO:3, wherein x represents an integer from 1 to 26, inclusive, and y represents an integer from 209 to 219, inclusive, of SEQ ID NO:3;  
 (iii) a polypeptide comprising amino acids x to y of SEQ ID NO:4, wherein x represents an integer from 1 to 26, inclusive, and y represents an integer from 209 to 219, inclusive, of SEQ ID NO:4;  
 (iv) a polypeptide comprising amino acids x to y of SEQ ID NO:5, wherein x represents an integer from 1 to 26, inclusive, and y represents an integer from 209 to 219, inclusive, of SEQ ID NO:5;  
 (v) a polypeptide comprising amino acids x to y of SEQ ID NO:6, wherein x represents an integer from 1 to 26, inclusive, and y represents an integer from 209 to 219, inclusive, of SEQ ID NO:6;  
 (vi) a polypeptide comprising amino acids x to y of SEQ ID NO:7, wherein x represents an integer from 1 to 26, inclusive, and y represents an integer from 208 to 218, inclusive, of SEQ ID NO:7;  
 (vii) a polypeptide comprising amino acids x to y of SEQ ID NO:8, wherein x represents an integer from 1 to 26, inclusive, and y represents an integer from 208 to 218, inclusive, of SEQ ID NO:8; and  
 (viii) a polypeptide comprising amino acids x to y of SEQ ID NO:9, wherein x represents an integer from 1 to 26, inclusive, and y represents an integer from 208 to 218, inclusive, of SEQ ID NO:9, wherein the A41L polypeptide binds LAR, and  
   (b) determining whether the test compound alters the ability of A41L to bind LAR.    
     
     
         6 . A method for identifying compounds that alter one or more biological activities of LAR, comprising: 
 (a) mixing a test compound with a LAR polypeptide selected from the group consisting of 
 (i) a polypeptide comprising amino acids x to y, wherein x represents an integer from 1 to 20, inclusive, and y represents an integer from 960 to 965, inclusive, of SEQ ID NO:10;  
 (ii) a polypeptide comprising amino acids x to y, wherein x represents an integer from 1 to 20, inclusive, and y represents an integer from 302 to 312, inclusive, of SEQ ID NO:10;  
 (iii) a polypeptide comprising amino acids x to y, wherein x represents an integer from 1 to 20, inclusive, and y represents an integer from 296 to 306, inclusive, of SEQ ID NO:12;  
 (iv) fragments of the polypeptides of (i) through (iii) that are capable of binding A41L; and  
 (v) fusion proteins comprising a polypeptide of (I) through (iv); and  
   (b) determining whether the test compound alters the ability of LAR to bind A41L.    
     
     
         7 . A peptide comprising amino acid 181 through 186 of SEQ ID NO:10.  
     
     
         8 . An antibody that binds the peptide of  claim 14 .  
     
     
         9 . The antibody of  claim 8  which is a monoclonal antibody.  
     
     
         10 . The antibody of  claim 8 , which does not bind other forms of LAR.  
     
     
         11 . The antibody of  claim 10  which is a monoclonal antibody.  
     
     
         12 . A polynucleotide comprising nucleotides 1 through 18 of SEQ ID NO:13.  
     
     
         13 . A method for altering one or more biological activities of LAR comprising contacting a cell expressing LAR with least one compound selected according to the method of  claim 5 .  
     
     
         14 . A method for altering one or more biological activities of LAR comprising contacting a cell expressing LAR with at least one compound selected according to the method of  claim 6 .  
     
     
         15 . A polypeptide selected from the group consisting of: 
 (a) a polypeptide comprising amino acids x to y, wherein x represents an integer from 1 to 20, inclusive, and y represents an integer from 960 to 965, inclusive, of SEQ ID NO:10;    (b) a polypeptide comprising amino acids x to y, wherein x represents an integer from 1 to 20, inclusive, and y represents an integer from 302 to 312, inclusive, of SEQ ID NO:10;    (c) a polypeptide comprising amino acids x to y, wherein x represents an integer from 1 to 20, inclusive, and y represents an integer from 296 to 306, inclusive, of SEQ ID NO:12; and    (d) a fragment of the polypeptides of (a), (b), or (c), wherein the fragment is capable of binding an A41L polypeptide.    
     
     
         16 . A fusion protein comprising the polypeptide of  claim 15 .  
     
     
         17 . The fusion protein of  claim 16 , wherein the peptide that promotes oligomerization is selected from the group consisting of a leucine zipper and an immunoglobulin Fc domain.