Influence of active site location on catalytic activity in de novo-designed zinc metalloenzymes.

ABSTRACT: While metalloprotein design has now yielded a number of successful metal-bound and even catalytically active constructs, the question of where to put a metal site along a linear, repetitive sequence has not been thoroughly addressed. Often several possibilities in a given sequence may exist that would appear equivalent but may in fact differ for metal affinity, substrate access, or protein dynamics. We present a systematic variation of active site location for a hydrolytically active ZnHis3O site contained within a de novo-designed three-stranded coiled coil. We find that the maximal rate, substrate access, and metal-binding affinity are dependent on the selected position, while catalytic efficiency for p-nitrophenyl acetate hydrolysis can be retained regardless of the location of the acti

SUBMITTER: Zastrow ML 

PROVIDER: S-EPMC3667658 | biostudies-literature | 2013 Apr

REPOSITORIES: biostudies-literature