Site-Specific Incorporation of a Thioester Containing Amino Acid into Proteins.
Weimin XuanDaniel CollinsMinseob KohSida ShaoAnzhi YaoHan XiaoPhilip GarnerPeter G SchultzPublished in: ACS chemical biology (2018)
Here, we report the site-specific incorporation of a thioester containing noncanonical amino acid (ncAA) into recombinantly expressed proteins. Specifically, we genetically encoded a thioester-activated aspartic acid (ThioD) in bacteria in good yield and with high fidelity using an orthogonal nonsense suppressor tRNA/aminoacyl-tRNA synthetase (aaRS) pair. To demonstrate the utility of ThioD, we used native chemical ligation to label green fluorescent protein with a fluorophore in good yield.