Premium
Multicolor time‐resolved Förster resonance energy transfer microscopy reveals the impact of GPCR oligomerization on internalization processes
Author(s) -
Faklaris Orestis,
Cottet Martin,
Falco Amandine,
Villier Brice,
Laget Michel,
Zwier Jurriaan M.,
Trinquet Eric,
Mouillac Bernard,
Pin JeanPhilippe,
Durroux Thierry
Publication year - 2015
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fj.14-260059
Subject(s) - internalization , förster resonance energy transfer , g protein coupled receptor , biophysics , oligomer , microscopy , chemistry , energy transfer , receptor , nanotechnology , biological system , microbiology and biotechnology , biology , materials science , physics , chemical physics , fluorescence , biochemistry , organic chemistry , quantum mechanics , optics
Identifying the interacting partners and the dynamics of the molecular networks constitutes the key point in understanding cellular processes. Different methods often based on energy transfer strategies have been developed to examine the molecular dynamics of protein complexes. However, these methods suffer a couple of drawbacks: a single complex can be studied at a time, and its localization and tracking cannot generally be investigated. Here, we report a multicolor time‐resolved Förster resonance energy transfer microscopy method that allows the identification of up to 3 different complexes simultaneously, their localization in cells, and their tracking after activation. Using this technique, we studied GPCR oligomerization and internalization in human embryonic kidney 293 cells. We definitively show that receptors can internalize as oligomers and that receptor coexpression deeply impacts oligomer internalization processes.— Faklaris, O., Cottet, M., Falco, A., Villier, B., Laget, M., Zwier, J. M., Trinquet, E., Mouillac, B., Pin, J.‐P., Durroux, T. Multicolor time‐resolved Förster resonance energy transfer microscopy reveals the impact of GPCR oligomerization on internalization processes. FASEB J. 29, 2235‐2246 (2015). www.fasebj.org