Patent ID: 6797482
Filing Date: 2004-09-28
Classification: G01N,Y10T

Abstract:
A method of characterizing a binding site of a binding protein of known or determinable amino acid sequence, said method comprising the steps of:(a) selectively labeling peptide amide groups in a binding site of the binding protein that have been determined to be substantially protected from exchange with solvent hydrogens in a situation where the binding protein binds its binding partner; (b) progressively degrading, under conditions of slow hydrogen exchange, the selectively labeled binding protein to obtain at least one series of subfragments, wherein each subfragment of each of said series is shorter than the preceding subfragment in the series by about 1-5 fewer amino acid residues at one terminus; (c) quantifying an amount of heavy hydrogen on each subfragment; and (d) correlating the amount of label on the subfragments with the amino acid sequence of the binding protein, thereby localizing the positions of the binding protein that had been labeled to within about 1-5 amino acid residues and thus characterizing the binding site of the binding protein.