Site-Specific Incorporation of a Thioester Containing Amino Acid into Proteins

[Image: see text] 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...

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Detalles Bibliográficos
Autores principales: Xuan, Weimin, Collins, Daniel, Koh, Minseob, Shao, Sida, Yao, Anzhi, Xiao, Han, Garner, Philip, Schultz, Peter G.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: American Chemical Society 2018
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5856652/
https://www.ncbi.nlm.nih.gov/pubmed/29360343
http://dx.doi.org/10.1021/acschembio.7b00998
Descripción
Sumario:[Image: see text] 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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