Design of activated serine-containing catalytic triads with atomic-level accuracy.

ABSTRACT: A challenge in the computational design of enzymes is that multiple properties, including substrate binding, transition state stabilization and product release, must be simultaneously optimized, and this has limited the absolute activity of successful designs. Here, we focus on a single critical property of many enzymes: the nucleophilicity of an active site residue that initiates catalysis. We design proteins with idealized serine-containing catalytic triads and assess their nucleophilicity directly in native biological systems using activity-based organophosphate probes. Crystal structures of the most successful designs show unprecedented agreement with computational models, including extensive hydrogen bonding networks between the catalytic triad (or quartet) residues, and mutagenesis e

SUBMITTER: Rajagopalan S 

PROVIDER: S-EPMC4048123 | biostudies-literature | 2014 May

REPOSITORIES: biostudies-literature