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Chemical-Tag Labeling of Proteins Using Fully Recombinant Split Inteins

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Site-Specific Protein Labeling

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1266))

Abstract

Chemical-tag labeling of proteins involving split inteins is an approach for the selective chemical modification of proteins without the requirement of any chemical synthesis to be performed. In a two-step protocol, a very short tag fused to a split intein auxiliary protein is first labeled in a bioconjugation reaction with a synthetic moiety either at its N-terminus (amine-tag) or at the side chain of an unnatural amino acid (click-tag). The labeled protein is then mixed with the protein of interest fused to the complementary intein fragment. In the resulting spontaneous protein trans-splicing reaction the split intein fragments remove themselves and ligate the tag to the protein of interest in a virtually traceless fashion. The reaction can be performed either using a purified protein of interest or to label a protein in the context of a living cell. All protein components are recombinantly expressed and all chemical reagents are commercially available.

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Acknowledgements

We thank Peter G. Schultz (The Scripps Research Institute) for providing plasmids for AzF incorporation. Financial support was kindly provided by the DFG (SPP1623).

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Authors and Affiliations

  1. Department of Chemistry and Pharmacy, Institute of Biochemistry, University of Muenster, Wilhelm-Klemm-Str. 2, 48149, Münster, Germany

    Anne-Lena Bachmann, Julian C. J. Matern, Vivien Schütz & Henning D. Mootz

Authors
  1. Anne-Lena Bachmann
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  2. Julian C. J. Matern
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  3. Vivien Schütz
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  4. Henning D. Mootz
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Corresponding author

Correspondence to Henning D. Mootz .

Editor information

Editors and Affiliations

  1. Department of Chemistry, École Normale Supérieure, Paris, France

    Arnaud Gautier

  2. Sensitive Farbstoffe GbR, Munich, Germany

    Marlon J. Hinner

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Bachmann, AL., Matern, J.C.J., Schütz, V., Mootz, H.D. (2015). Chemical-Tag Labeling of Proteins Using Fully Recombinant Split Inteins. In: Gautier, A., Hinner, M. (eds) Site-Specific Protein Labeling. Methods in Molecular Biology, vol 1266. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-2272-7_10

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  • DOI: https://doi.org/10.1007/978-1-4939-2272-7_10

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-2271-0

  • Online ISBN: 978-1-4939-2272-7

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