A Single Mutation in a Regulatory Protein Produces Evolvable Allosterically Regulated Catalyst of Nonnatural Reaction

• DOI: 10.1002/ange.201302339
• Publication Date: 2013-04-29T19:44:28Z
• Authors: Olesia V. Moroz; Yurii S. Moroz; Yibing Wu; Alissa B. Olsen; Hong Cheng; Korrie L. Mack; Jaclyn M. McLaughlin; Elizabeth A. Raymond; Krystyna Zhezherya; Heinrich Roder; Ivan V. Korendovych
• Keywords: Models, Molecular; 0301 basic medicine; 0303 health sciences; 03 medical and health sciences; Allosteric Regulation; Mutation; Proteins; Catalysis

It only takes one mutation: a strategically placed single mutation in a non-enzymatic protein scaffold produced AlleyCat, a small, allosterically regulated catalyst of Kemp elimination. In only 7 rounds of directed evolution enzymatic efficiency of the original 74 amino acid residue catalyst was improved more than 220-fold to achieve kcat value higher than that of catalytic antibodies for the same reaction, still preserving allosteric regulation.