Methods of Identifying Modulators of Interaction Between PDZ Domain Proteins and PL Proteins

Abstract

The invention provides reagents and methods for inhibiting or enhancing interactions between proteins in hematopoietic cells and other cells involved in the mediation of an immune response. Reagents and methods provided are useful for treatment of a variety of diseases and conditions mediated by immune system cells.

1 - 48 . (canceled) 
     
     
         49 . A method of determining whether a test compound is an inhibitor or agonist of binding between a PDZ protein and a PL protein, comprising:
 (a) contacting a peptide comprising a PDZ domain (PDZ) with a plurality of PL peptides (PL) in the presence and in the absence of a test compound,   (b) detecting any interaction between the PDZ and at least one PL in the presence and absence of the test compound,   wherein   (i) less interaction between the PDZ and at least one PL in the presence of the test compound than in the absence of the compound indicates that the test compound is an inhibitor of such an interaction, or   (ii) more interaction between the PDZ and at least one PL in the presence of the test compound than in the absence of the compound indicates that the test compound is an agonist of such an interaction.   
     
     
         50 . The method of  claim 49 , wherein the PDZ is contacted with the plurality of PLs simultaneously. 
     
     
         51 . The method of  claim 50 , wherein the PLs are organized in an array. 
     
     
         52 . The method of  claim 51 , wherein the different PLs are situated in separate areas of the array. 
     
     
         53 . The method of  claim 51 , wherein the different PLs are each situated in a different well of a multi-well plate. 
     
     
         54 . The method of  claim 51 , wherein the PL peptides are arrayed within a common area of a surface such that they can be simultaneously exposed to the same solution. 
     
     
         55 . The method of  claim 51 , wherein the method comprises the use of multiple arrays of PLs in tandem. 
     
     
         56 . The method of  claim 49 , wherein at least one of the plurality of PLs is immobilized. 
     
     
         57 . The method of  claim 56 , wherein the at least one of the plurality of PLs is bound to the surface of an assay plate. 
     
     
         58 . The method of  claim 57 , wherein the assay plate is a multi-well plate. 
     
     
         59 . The method of  claim 57 , wherein the assay plate is a high density array. 
     
     
         60 . The method of  claim 49 , wherein the test agent comprises a peptide having at its carboxy-terminus at least two amino acids that are the same as the carboxy-terminal two residues of a PL protein. 
     
     
         61 . The method of  claim 60 , wherein the test agent comprises a motif X-S-X-A, X-A-D/E-V, X-V/I/L-X*-V, or X-S/T-X-F, (where X is any amino acid and X* is any non-aromatic amino acid). 
     
     
         62 . The method of  claim 49 , wherein at least one of the plurality of PLs is bound to a substrate in the form of a distinguishable bead. 
     
     
         63 . The method of  claim 49 , further comprising determining the effect on the test agent upon a plurality of PDZ-PL pairs. 
     
     
         64 . The method of  claim 63 , wherein the determining is performed simultaneously on the plurality of PDZ-PL pairs. 
     
     
         65 . The method of  claim 64 , further comprising selecting a test agent that modulates the binding of a first PDZ-PL pair of interest, but does not inhibit, or inhibits to a lesser extent, the binding of a second different PDZ-PL pair of interest. 
     
     
         66 . The method of  claim 65 , wherein the first PDZ-PL pair comprises the same PDZ as the second PDZ-PL pair, but comprises a different PL. 
     
     
         67 . The method of  claim 65 , wherein the first PDZ-PL pair comprises a different PDZ and a different PL from the second PDZ-PL pair. 
     
     
         68 . The method of  claim 49 , wherein the method involves determining whether a test compound is an inhibitor.