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

  • Xi Chen
  • Fu Li
  • Yao-Wen WuEmail author
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 2008)

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.

Key words

Protein labeling Chemical probes In vivo chemical labeling eDHFR tag Affinity conjugation Bioorthogonal reactions 

Notes

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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Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2019

Authors and Affiliations

  1. 1.Chemical Genomics Centre of the Max Planck SocietyDortmundGermany
  2. 2.Max Planck Institute for Molecular PhysiologyDortmundGermany
  3. 3.Department of ChemistryUmeå UniversityUmeåSweden

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