Discovery of Escherichia coli methionyl-tRNA synthetase mutants for efficient labeling of proteins with azidonorleucine in vivo
I. Caglar Tanrikulu, Emmanuelle Schmitt, Yves Mechulam, William A. Goddard III, David A. Tirrell
Abstract
Incorporation of noncanonical amino acids into cellular proteins often requires engineering new aminoacyl-tRNA synthetase activity into the cell. A screening strategy that relies on cell-surface display of reactive amino acid side-chains was used to identify a diverse set of methionyl-tRNA synthetase (MetRS) mutants that allow efficient incorporation of the methionine (Met) analog azidonorleucine (Anl). We demonstrate that the extent of cell-surface labeling in vivo is a good indicator of the rate of Anl activation by the MetRS variant harbored by the cell. By screening at low Anl concentrations in Met-supplemented media, MetRS variants with improved activities toward Anl and better discrimination against Met were identified.
Group Members
Tanrikulu, I. C., Schmitt, E., Mechulam, Y., III, W. A. G., & Tirrell, D. A. (2009). Discovery of Escherichia coli methionyl-tRNA synthetase mutants for efficient labeling of proteins with azidonorleucine in vivo. *Proc. Natl. Acad. Sci. U.S.A.*, *106*(36), 15285-15290. https://doi.org/10.1073/pnas.0905735106