Fluorogenic Probing of Membrane Protein Trafficking

Chenge Li (1, 2) , Aurélien Mourton (3, 2) , Marie-Aude Plamont (1, 2) , Vanessa Rodrigues (1, 2) , Isabelle Aujard (1) , Michel Volovitch (3, 4, 2) , Thomas Le Saux (1) , Franck Perez (5, 2) , Sophie Vriz (3, 6) , Ludovic Jullien (1) , Alain Joliot (3) , Arnaud Gautier (1, 2)

1 PASTEUR - Processus d'Activation Sélective par Transfert d'Energie Uni-électronique ou Radiatif (UMR 8640)
2 PSL - Université Paris Sciences et Lettres
3 CIRB - Centre interdisciplinaire de recherche en biologie
4 Département de Biologie - ENS Paris
5 Institut Curie [Paris]
6 UPD7 - Université Paris Diderot - Paris 7

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.

Domaines: Chimie organique; Biochimie, Biologie Moléculaire

Chenge Li, Aurélien Mourton, Marie-Aude Plamont, Vanessa Rodrigues, Isabelle Aujard, et al.. Fluorogenic Probing of Membrane Protein Trafficking. Bioconjugate Chemistry, 2018, 29 (6), pp.1823-1828. ⟨10.1021/acs.bioconjchem.8b00180⟩. ⟨hal-01919676⟩