Conformational Ensembles Reveal the Origins of Serine Protease Catalysis

Abstract

AbstractEnzymes exist in ensembles of states that encode the energetics underlying their catalysis. Conformational ensembles built from 1231 structures of 17 serine proteases reveal atomic-level changes across their reaction states, identify molecular features that provide catalysis, and quantify their energetic contributions to catalysis. These enzymes precisely position their reactants in destabilized conformers, creating a downhill energetic gradient that selectively favors the motions required for reaction while limiting off-pathway conformational states. A local catalytic motif, the “nucleophilic elbow”, has repeatedly evolved, generating ground state destabilization in 50 proteases and 52 additional enzymes spanning 32 distinct structural folds. Ensemble–function analyses reveal previously unknown catalytic features, provide quantitative models based on simple physical and chemical principles, and identify motifs recurrent in Nature that may inspire enzyme design.One sentence summary:Ensemble–function analyses provide a quantitative model for serine protease catalysis, reveal previously unknown conformational features that contribute to their catalysis, and identify a structural motif that underlie these features and has evolved in >100 different enzymes from 32 protein folds.