Patent ID: 6797460
Filing Date: 2004-09-28
Classification: C12Q,G01N,Y10T

Abstract:
A method for identifying an atom of a common ligand mimic that is proximal to an interface region of an enzyme;wherein the enzyme can bind a common ligand (CL) or a common ligand mimic (CL mimic) at a common ligand site (CL site) and can bind a specificity ligand (SL) at an adjacent specificity ligand site (SL site); wherein an interface region is defined as the atoms of the enzyme between the CL site and SL site, and atoms of an SL, if bound to the enzyme; wherein the enzyme can catalyze a reaction involving the SL and a reactive atom of the CL; and wherein a CL reactive region is defined as CL atoms immediately adjacent to the SL; comprising the steps of: (a) identifying an atom of the interface region, comprising the steps of: (1) binding an SL to the SL site of said enzyme; (2) irradiating a nucleus of an atom of the SL reactive region; and (3) identifying in a multidimensional NMR experiment an NMR cross-peak corresponding to a nucleus of said enzyme that is perturbed by the irradiation of the nucleus of the SL reactive region, thereby identifying an atom of the interface region; then (b) identifying an atom in a CL mimic that is proximal to the interface region, comprising the steps of: (1) binding a CL mimic to the CL site of said enzyme; (2) irradiating the nucleus of the interface atom identified in step (a); and (3) identifying in a multidimensional NMR experiment an NMR cross-peak corresponding to a nucleus of the CL mimic that is perturbed by the irradiation of the interface nucleus, thereby identifying an atom of the CL mimic that is proximal to the interface region.