METHOD FOR IDENTIFYING BINDING SITE OR ITS VICINITY OF SUBSTRATE OR ITS ANALOG AND/OR ALLOSTERIC LIGAND IN ALLOSTERIC PROTEIN, METHOD FOR DETERMINING ORDER OF BINDING LIGAND TO ALLOSTERIC PROTEIN, AND METHOD FOR DETECTING ALLOSTERIC LIGAND

摘要

PROBLEM TO BE SOLVED: To provide a technique for analyzing structural change by binding between an allosteric ligand and a substrate in an allosteric protein with a minute amount of a sample in a short time. SOLUTION: The subject method for identifying the binding site or its vicinity of a substrate or its analog and/or an allosteric ligand in the allosteric protein comprises (i) the step of deuterating an allosteric protein, (ii) the step of measuring the mass of the deuterated protein, (iii) the step of obtaining a peptide mixture by fragmenting the deuterated protein, (iv) the step of measuring the respective masses of the peptides in the peptide mixture, (v) the step of calculating the deuteration percentage of each of the fragmented peptides, (vi) the step of calculating the relative deuteration percentage for the respective fragmented peptides and (vii) the step of identifying a peptide containing the allosteric ligand binding site and/or a peptide containing the substrate or its analog binding site or its vicinity based on the relative deuteration percentage for the respective fragmented peptides. COPYRIGHT: (C)2006,JPO&NCIPI