A single mutation in a regulatory protein produces evolvable allosterically regulated catalyst of nonnatural reaction
Angew Chem Int Ed Engl. 2013 Jun 10;52(24):6246-9.
doi: 10.1002/anie.201302339.
Epub 2013 Apr 29.
- PMID: 23630096
- PMCID: PMC3817844
- DOI: 10.1002/anie.201302339
Abstract
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.
MeSH terms
- Allosteric Regulation
- Catalysis
- Models, Molecular
- Mutation*
- Proteins / chemistry
- Proteins / genetics*
- Proteins / metabolism
Substances
- Proteins