Site-Specific Incorporation of a Thioester Containing Amino Acid into Proteins
Weimin Xuan1, Daniel Collins2, Minseob Koh1
1Department of Chemistry and Skaggs Institute for Chemical Biology , The Scripps Research Institute , 10550 N Torrey Pines Road , La Jolla , California 92037 , United States.
Abstract:
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.
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