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Bioconjug Chem. 2018 Jun 20;29(6):1823-1828. doi: 10.1021/acs.bioconjchem.8b00180. Epub 2018 May 25.

Fluorogenic Probing of Membrane Protein Trafficking.

Author information

1
PASTEUR, Département de Chimie, École Normale Supérieure , PSL University, Sorbonne Université, CNRS , 75005 Paris , France.
2
Center for Interdisciplinary Research in Biology (CIRB), Collège de France , CNRS, INSERM, PSL Research University , 75231 Paris , France.
3
PSL Research University , 75006 Paris , France.
4
École Normale Supérieure, Department of Biology , PSL Research University , 75005 Paris , France.
5
Institut Curie, PSL Research University, CNRS UMR144 , 26 rue d'Ulm , 75248 Paris Cedex 05, France.
6
Université Paris Diderot, Sorbonne Paris Cité , Biology Department , 75205 Paris Cedex 13, France.

Abstract

Methods to differentially label cell-surface and intracellular membrane proteins are indispensable for understanding their function and the regulation of their trafficking. We present an efficient strategy for the rapid and selective fluorescent labeling of membrane proteins based on the chemical-genetic fluorescent marker FAST (fluorescence-activating and absorption-shifting tag). Cell-surface FAST-tagged proteins could be selectively and rapidly labeled using fluorogenic membrane-impermeant 4-hydroxybenzylidene rhodanine (HBR) analogs. This approach allows the study of protein trafficking at the plasma membrane with various fluorometric techniques, and opens exciting prospects for the high-throughput screening of small molecules able to restore disease-related trafficking defects.

PMID:
29791141
DOI:
10.1021/acs.bioconjchem.8b00180
[Indexed for MEDLINE]

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