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

Amit Sachdeva, Kaihang Wang, Thomas Elliott, Jason W. Chin

Research output: Contribution to journalArticlepeer-review

123 Citations (Scopus)

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.
Original languageEnglish
Pages (from-to)7785-7788
Number of pages4
JournalJournal of the American Chemical Society
Volume136
Issue number22
DOIs
Publication statusPublished - 4 Jun 2014

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