Concerted, rapid, quantitative, and site-specific dual labeling of proteins

Sachdeva, Amit, Wang, Kaihang, Elliott, Thomas and Chin, Jason W. (2014) Concerted, rapid, quantitative, and site-specific dual labeling of proteins. Journal of the American Chemical Society, 136 (22). pp. 7785-7788. ISSN 0002-7863

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Abstract

Rapid, one-pot, concerted, site-specific labeling of proteins at genetically encoded unnatural amino acids with distinct small molecules at physiological pH, temperature, and pressure is an important challenge. Current approaches require sequential labeling, low pH, and typically days to reach completion, limiting their utility. We report the efficient, genetically encoded incorporation of alkyne- and cyclopropene-containing amino acids at distinct sites in a protein using an optimized orthogonal translation system in E. coli. and quantitative, site-specific, one-pot, concerted protein labeling with fluorophores bearing azide and tetrazine groups, respectively. Protein double labeling in aqueous buffer at physiological pH, temperature, and pressure is quantitative in 30 min.

Item Type:	Article
Faculty \ School:	Faculty of Science > School of Chemistry (former - to 2024)
UEA Research Groups:	Faculty of Science > Research Groups > Chemistry of Life Processes
Depositing User:	Pure Connector
Date Deposited:	24 Jul 2015 22:38
Last Modified:	28 Mar 2025 06:37
URI:	https://ueaeprints.uea.ac.uk/id/eprint/53628
DOI:	10.1021/ja4129789

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