Probing trypsin specificity with basic 3-substituted glutarate diesters and related monoester substrate analogs: Evidence for allosteric activation of trypsin catalysis

Abstract

A series of dimethyl glutarates bearing basic substituents at C-3, and related monoesters, have been evaluated as substrates of trypsin in order to probe the asymmetric synthetic potential of the enzyme with respect to enantiotopic ester group, and enantiomer, discrimination. While none of the mono- or diesters proved to be a trypsin substrate, several of them accelerated trypsin-catalyzed hydrolysis of the standard reference substrate BAEE, in a manner consistent with an allosteric activation process. The results provide the first examples of allosteric activation of trypsin by modifiers that are sterically precluded from interacting effectively at the acive site.