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Ligand-directed tosyl chemistry for protein labeling in vivo

Nature Chemical Biology volume 5, pages 341–343 (2009)Cite this article

Abstract

Here we describe a method for the site-selective attachment of synthetic molecules into specific 'endogenous' proteins in vivo using ligand-directed tosyl (LDT) chemistry. This approach was applied not only for chemically labeling proteins in living cells, tissues and mice but also for constructing a biosensor directly inside cells without genetic engineering. These data establish LDT chemistry as a new tool for the study and manipulation of biological systems.

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Figure 1: LDT chemistry for labeling endogenous proteins in living native cells.
Figure 2: CA-selective labeling in vitro and in vivo.
Figure 3: Functional conversion of endogenous CA into a 19F NMR–based biosensor in RBCs.

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Acknowledgements

We thank M. Shirakawa and H. Tochio (Kyoto University) for helpful discussions and assistance with 19F NMR measurements. We are grateful to Y. Mori and all members of his group (Kyoto University), in particular S. Kiyonaka, Y. Uriu and S. Yamamoto, for generous assistance on animal experiments. We thank E. Ashihara (Kyoto University Hospital) for his help in taking blood samples. We thank M. Yamada (Shimadzu Corporation) for measuring tandem mass spectra (MALDI-QIT-TOF MS/MS). We also thank all members of the Hamachi laboratory, particularly H. Nonaka, for discussions and experimental assistance. Y.T. acknowledges the Japan Society for the Promotion of Science Research Fellowships for Young Scientists.

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Authors and Affiliations

  1. Department of Synthetic Chemistry and Biological Chemistry, Graduate School of Engineering, Kyoto University, Katsura, Nishikyo-ku, Kyoto, Japan

    Shinya Tsukiji, Masayoshi Miyagawa, Yousuke Takaoka, Tomonori Tamura & Itaru Hamachi

  2. Core Research for Evolutional Science and Technology (CREST), Japan Science and Technology Agency, Sanbancho, Chiyoda-ku, Tokyo, Japan

    Itaru Hamachi

Authors
  1. Shinya Tsukiji
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  2. Masayoshi Miyagawa
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  3. Yousuke Takaoka
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  4. Tomonori Tamura
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Contributions

S.T. and I.H. conceived and designed the project. S.T. performed the eCA labeling in mice. M.M. performed the synthesis and the in vitro CAII labeling. Y.T. performed the synthesis, the eCA labeling in RBCs, including the 19F NMR measurements, and the congerin labeling. T.T. performed the synthesis and the FKBP12 labeling. The manuscript was written by S.T. and I.H.

Corresponding author

Correspondence to Itaru Hamachi.

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Supplementary Figures 1–7, Supplementary Table 1 and Supplementary Methods (PDF 4820 kb)

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Tsukiji, S., Miyagawa, M., Takaoka, Y. et al. Ligand-directed tosyl chemistry for protein labeling in vivo. Nat Chem Biol 5, 341–343 (2009). https://doi.org/10.1038/nchembio.157

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