Journal of the American Chemical Society · 2006 · 103 citations · 6 references
Life SciencesBiochemistryBiotechnologyBiocatalysisEnzymologyProtein EngineeringActive Site EngineeringEnzyme ActivatorsEnzyme Active SitesEnzyme DesignEnzyme FunctionProtein Engineering Of EnzymesActive Site DesignActive SitesAmino AcidsEnzyme Active Site ArchitectureEnzyme Active Site DesignEnzyme Active Site EngineeringNitroreductase-activated ProdrugsUnnatural Amino Acid Incorporation
The ability to site-specifically incorporate a diverse set of unnatural amino acids (>30) into proteins and quickly add new structures of interest has recently changed our approach to protein use and study. One important question yet unaddressed with unnatural amino acids (UAAs) is whether they can improve the activity of an enzyme beyond that available from the natural 20 amino acids. Herein, we report the >30-fold improvement of prodrug activator nitroreductase activity with an UAA over that of the native active site and a >2.3-fold improvement over the best possible natural amino acid. Because immense structural and electrostatic diversity at a single location can be sampled very quickly, UAAs can be implemented to improve enzyme active sites and tune a site to multiple substrates.
6