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Genetically Encoded Cyclopropene Directs Rapid, Photoclick-Chemistry-Mediated Protein Labeling in Mammalian Cells

Authors

  • Dr. Zhipeng Yu,

    1. Department of Chemistry, State University of New York at Buffalo, Buffalo, NY 14260 (USA)
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    • These two authors contributed equally to the work.

  • Yanchao Pan,

    1. Laboratory of Noncoding RNA, Institute of Biophysics, Chinese Academy of Sciences, Beijing 100101 (P.R. China)
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    • These two authors contributed equally to the work.

  • Dr. Zhiyong Wang,

    1. Department of Chemistry, State University of New York at Buffalo, Buffalo, NY 14260 (USA)
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  • Prof. Dr. Jiangyun Wang,

    Corresponding author
    1. Laboratory of Noncoding RNA, Institute of Biophysics, Chinese Academy of Sciences, Beijing 100101 (P.R. China)
    • Laboratory of Noncoding RNA, Institute of Biophysics, Chinese Academy of Sciences, Beijing 100101 (P.R. China)
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  • Prof. Dr. Qing Lin

    Corresponding author
    1. Department of Chemistry, State University of New York at Buffalo, Buffalo, NY 14260 (USA)
    • Department of Chemistry, State University of New York at Buffalo, Buffalo, NY 14260 (USA)
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  • We gratefully acknowledge the National Institutes of Health (GM 085092 to Q.L.), the Major State Basic Research Program of China (2010CB912301 and 2009CB825505 to J.W.), National Science Foundation of China (90913022 to J.W.), and CAS (KZCX2-YW-JC101 and KSCX2-EW-G-7 to J.W.) for financial support. We thank William Brennessel at the University of Rochester for X-ray crystallography and Reyna K. Lim in the Q.L. lab for plasmid preparation, Alan Siegel at SUNY Buffalo Biological Sciences Imaging Facility (supported by National Science Foundation Major Research Instrumentation grant DBI-0923133) for assistance in microscopy. CCDC 808108 (1 c) contains the supplementary crystallographic data for this paper. These data can be obtained free of charge from The Cambridge Crystallographic Data Centre via www.ccdc.cam.ac.uk/data_request/cif.

Abstract

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We just click: Genetic incorporation of a cyclopropene amino acid CpK (see scheme) site-specifically into proteins in E. coli and mammalian cells was achieved using an orthogonal aminoacyl-tRNA synthetase/tRNACUA pair (CpKRS/MbtRNACUA). Cyclopropene exhibited fast reaction kinetics in the photoclick reaction and allowed rapid (ca. 2 min) labeling of proteins.

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