Transient Formation of a Complex Between α-Chymotrypsin, Proflavin, and Tosylarginine Methyl Ester (TAME)

Abstract

Chemical relaxation experiments were conducted on the reaction of α-chymotrypsin, with the competitive inhibitor proflavin and the substrate analogue TAME (tosylarginine methyl ester) in phosphate buffer, pH 6.7, observing transmission changes at 465 mμ. Two chemical relaxation processes were observed with the slow one attributed to a monomolecular interconversion of the enzyme–substrate complex. The concentration dependence of the reciprocal fast relaxation time constant only agrees with the equations derived for the involvement of a labile ternary complex between enzyme, substrate, and inhibitor (as simplest model).