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

Sachdeva, Amit and Wang, Kaihang and 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. PMCID PMC4333588. http://resolver.caltech.edu/CaltechAUTHORS:20180626-164508438

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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
Related URLs:
URLURL TypeDescription
https://doi.org/10.1021/ja4129789DOIArticle
https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4333588/PubMed CentralArticle
https://pubs.acs.org/doi/suppl/10.1021/ja4129789PublisherSupporting Information
Additional Information:© 2014 American Chemical Society. ACS AuthorChoice - This is an open access article published under a Creative Commons Attribution (CC-BY) License, which permits unrestricted use, distribution and reproduction in any medium, provided the author and source are cited. Received: December 20, 2013. Published: May 23, 2014. We are grateful to Medical Research Council (U105181009, UD99999908) and European Research Council (MC-A024-5PG0A) for funding this work. Author Contributions: A.S. and K.W. contributed equally. The authors declare no competing financial interest.
Funders:
Funding AgencyGrant Number
Medical Research Council (UK)U105181009
Medical Research Council (UK)UD99999908
European Research Council (ERC)MC-A024-5PG0A
PubMed Central ID:PMC4333588
Record Number:CaltechAUTHORS:20180626-164508438
Persistent URL:http://resolver.caltech.edu/CaltechAUTHORS:20180626-164508438
Official Citation:Concerted, Rapid, Quantitative, and Site-Specific Dual Labeling of Proteins Amit Sachdeva, Kaihang Wang, Thomas Elliott, and Jason W. Chin Journal of the American Chemical Society 2014 136 (22), 7785-7788 DOI: 10.1021/ja4129789
Usage Policy:No commercial reproduction, distribution, display or performance rights in this work are provided.
ID Code:87367
Collection:CaltechAUTHORS
Deposited By: George Porter
Deposited On:27 Jun 2018 14:45
Last Modified:27 Jun 2018 14:45

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