Patent ID: 6620589
Filing Date: 2003-09-16
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; wherein a CL reactive region is defined as the reactive atom of the CL and CL atoms immediately adjacent to the reactive atom or CL atoms immediately adjacent to the SL; comprising the steps of (1) mixing a CL and CL mimic with said enzyme to establish an equilibrium of a CL-enzyme complex and CL mimic-enzyme complex; (2) irradiating a nucleus of an atom of the CL, whereby energy is transferred from the nucleus of the CL atom to the interface region; (3) observing a perturbation of the nucleus of an atom of a CL mimic, whereby energy is transferred from the interface region to said atom of the CL, mimic; and (4) identifying in a multidimensional NMR experiment an NMR cross-peak corresponding to a nucleus of the CL mimic perturbed in step (3), thereby identifying an atom of the CL mimic that is proximal to the interface region.