A general method for the covalent labeling of fusion proteins with small molecules in vivo (original) (raw)

Nature Biotechnology volume 21, pages 86–89 (2003)Cite this article

Abstract

Characterizing the movement, interactions, and chemical microenvironment of a protein inside the living cell is crucial to a detailed understanding of its function. Most strategies aimed at realizing this objective are based on genetically fusing the protein of interest to a reporter protein that monitors changes in the environment of the coupled protein. Examples include fusions with fluorescent proteins, the yeast two-hybrid system, and split ubiquitin1,2,3. However, these techniques have various limitations, and considerable effort is being devoted to specific labeling of proteins in vivo with small synthetic molecules capable of probing and modulating their function. These approaches are currently based on the noncovalent binding of a small molecule to a protein, the formation of stable complexes between biarsenical compounds and peptides containing cysteines, or the use of biotin acceptor domains4,5,6,7,8,9,10. Here we describe a general method for the covalent labeling of fusion proteins in vivo that complements existing methods for noncovalent labeling of proteins and that may open up new ways of studying proteins in living cells.

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Acknowledgements

Funding of this work was provided by the Swiss Science Foundation, the European Community, and the EPFL. S.G. was supported by a fellowship from the Boehringer Ingelheim Foundation. Nils Johnsson is acknowledged for helpful discussions and advice, Maik Kindermann for help with the synthesis of BGBT, Stefan Pitsch for the gift of 6-chloroguanine, Robert Damoiseaux for the synthesis of oligonucleotide containing BGBT, and Bernd Kaina for the CHO cell line.

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Author notes

  1. Antje Keppler and Susanne Gendreizig: These authors contributed equally to this work.

Authors and Affiliations

  1. Institute of Molecular and Biological Chemistry, Swiss Federal Institute of Technology Lausanne (EPFL), Lausanne, CH-1015, Switzerland
    Antje Keppler, Susanne Gendreizig, Thomas Gronemeyer & Kai Johnsson
  2. Institute of Biomolecular Sciences, Swiss Federal Institute of Technology Lausanne (EPFL), Lausanne, CH-1015, Switzerland
    Antje Keppler, Susanne Gendreizig, Thomas Gronemeyer, Horst Pick, Horst Vogel & Kai Johnsson

Authors

  1. Antje Keppler
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  2. Susanne Gendreizig
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  3. Thomas Gronemeyer
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  4. Horst Pick
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  5. Horst Vogel
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  6. Kai Johnsson
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Corresponding author

Correspondence toKai Johnsson.

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The authors declare no competing financial interests.

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Keppler, A., Gendreizig, S., Gronemeyer, T. et al. A general method for the covalent labeling of fusion proteins with small molecules in vivo.Nat Biotechnol 21, 86–89 (2003). https://doi.org/10.1038/nbt765

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