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Affinity Conjugation for Rapid and Covalent Labeling of Proteins in Live Cells

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Proximity Labeling

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2008))

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Abstract

Protein labeling is enormously useful for characterization of protein function in live cells and study of the related cellular processes. Covalent labeling of protein using affinity conjugation confers stable and selective labeling of protein in cells. Affinity conjugation combines a specific ligand-protein interaction with a proximity-induced reaction to selectively label the protein of interest (POI) in the cell. Therefore, either a fluorogenic probe is directly introduced to the POI or a bioorthogonal group is incorporated to the POI, which is subsequently labeled with a fluorescent probe. Here, we describe a method for affinity conjugation of protein with a fluorogenic probe and a “tagging-then-labeling” approach by a combination of affinity conjugation with bioorthogonal reactions.

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Acknowledgments

This work was supported by the Deutsche Forschungsgemeinschaft, DFG (grant No.: SPP 1623), European Research Council, ERC (ChemBioAP), Vetenskapsrådet (Nr. 2018-04585) and The Knut and Alice Wallenberg Foundation.

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Authors and Affiliations

  1. Chemical Genomics Centre of the Max Planck Society, Otto-Hahn-Str. 15, Dortmund, 44227, Germany

    Xi Chen, Fu Li & Yao-Wen Wu

  2. Max Planck Institute for Molecular Physiology, Dortmund, 44227, Otto-Hahn-Str. 11, Germany

    Xi Chen, Fu Li & Yao-Wen Wu

  3. Department of Chemistry, Umeå University, Umeå, 90187, Sweden

    Yao-Wen Wu

Authors
  1. Xi Chen
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  2. Fu Li
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  3. Yao-Wen Wu
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Corresponding author

Correspondence to Yao-Wen Wu .

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Editors and Affiliations

  1. Institute of Pharmacy and Molecular Biotechnology, Heidelberg University, Heidelberg, Germany

    Murat Sunbul  & Andres Jäschke  & 

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Chen, X., Li, F., Wu, YW. (2019). Affinity Conjugation for Rapid and Covalent Labeling of Proteins in Live Cells. In: Sunbul, M., Jäschke, A. (eds) Proximity Labeling. Methods in Molecular Biology, vol 2008. Humana, New York, NY. https://doi.org/10.1007/978-1-4939-9537-0_15

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  • DOI: https://doi.org/10.1007/978-1-4939-9537-0_15

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  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-4939-9536-3

  • Online ISBN: 978-1-4939-9537-0

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