New Tricks for Old Proteins: Single Mutations in a Nonenzymatic Protein Give Rise to Various Enzymatic Activities.

TitleNew Tricks for Old Proteins: Single Mutations in a Nonenzymatic Protein Give Rise to Various Enzymatic Activities.
Publication TypeJournal Article
Year of Publication2015
AuthorsMoroz, Y. S., T. T. Dunston, O. V. Makhlynets, O. V. Moroz, Y. Wu, J. H. Yoon, A. B. Olsen, J. M. McLaughlin, K. L. Mack, P. M. Gosavi, N. A. J. van Nuland, and I. V. Korendovych
JournalJ Am Chem Soc
Volume137
Issue47
Pagination14905-11
Date Published2015 Dec 02
ISSN1520-5126
KeywordsCatalysis, Circular Dichroism, Enzymes, Kinetics, Mutation, Nuclear Magnetic Resonance, Biomolecular, Proteins, Proteolysis, Spectrophotometry, Ultraviolet
Abstract

Design of a new catalytic function in proteins, apart from its inherent practical value, is important for fundamental understanding of enzymatic activity. Using a computationally inexpensive, minimalistic approach that focuses on introducing a single highly reactive residue into proteins to achieve catalysis we converted a 74-residue-long C-terminal domain of calmodulin into an efficient esterase. The catalytic efficiency of the resulting stereoselective, allosterically regulated catalyst, nicknamed AlleyCatE, is higher than that of any previously reported de novo designed esterases. The simplicity of our design protocol should complement and expand the capabilities of current state-of-art approaches to protein design. These results show that even a small nonenzymatic protein can efficiently attain catalytic activities in various reactions (Kemp elimination, ester hydrolysis, retroaldol reaction) as a result of a single mutation. In other words, proteins can be just one mutation away from becoming entry points for subsequent evolution.

DOI10.1021/jacs.5b07812
Alternate JournalJ. Am. Chem. Soc.
PubMed ID26555770
Research group: