Netrin receptors

Abstract

The invention provides methods and compositions relating to vertebrate UNC-5 proteins which function as receptor proteins for netrins, a family of cell guidance proteins. The proteins may be produced recombinantly from transformed host cells from the disclosed vertebrate UNC-5 encoding nucleic acid or purified from human cells. The invention provides specific hybridization probes and primers capable of specifically hybridizing with the disclosed vertebrate unc-5 gene, vertebrate UNC-5-specific binding agents such as specific antibodies, and methods of making and using the subject compositions in diagnosis, therapy and in the biopharmaceutical industry.

1 . An isolated vertebrate UNC-5 protein comprising SEQ ID NO: 5, 6, 7 or, 8, or a fragment thereof having vertebrate UNC-5-specific activity.  
     
     
         2 . An isolated protein according to  claim 1 , wherein said protein specifically binds a natural netrin protein.  
     
     
         3 . A recombinant nucleic acid encoding a protein according to  claim 1 .  
     
     
         4 . A cell comprising a nucleic acid according to  claim 3 .  
     
     
         5 . A method of making an isolated vertebrate UNC-5 protein, comprising steps: introducing a nucleic acid according to  claim 3  into a host cell or cellular extract, incubating said host cell or extract under conditions whereby said nucleic acid is expressed as a transcript and said transcript is expressed as a translation product comprising said protein, and isolating said translation product.  
     
     
         6 . An isolated vertebrate UNC-5 protein made by the method of  claim 5 .  
     
     
         7 . An isolated vertebrate unc-5 nucleic acid comprising SEQ ID NO: 1 ,2, 3, or 4, or a fragment thereof having at least 24 consecutive bases of SEQ ID NO: 1, 2, 3, or 4 and sufficient to specifically hybridize with a nucleic acid having the sequence of the corresponding SEQ ID NO: 1, 2, 3, or 4 in the presence of natural C. elegans unc-5 cDNA.  
     
     
         8 . A method of screening for an agent which modulates the binding of a vertebrate UNC-5 protein to a binding target, said method comprising the steps of: 
 incubating a mixture comprising: 
 an isolated protein according to  claim 1 ,  
 a binding target of said protein, and  
 a candidate agent;  
   under conditions whereby, but for the presence of said agent, said protein specifically binds said binding target at a reference affinity;    detecting the binding affinity of said protein to said binding target to determine an agent-biased affinity,    wherein a difference between the agent-biased affinity and the reference affinity indicates that said agent modulates the binding of said protein to said binding target.    
     
     
         9 . A method according to  claim 8 , wherein said binding target is a natural netrin protein.