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Improved donor/acceptor BRET couples for monitoring β‐arrestin recruitment to G protein‐coupled receptors
Author(s) -
Kamal Maud,
Marquez Marcel,
Vauthier Virginie,
Leloire Audrey,
Froguel Philippe,
Jockers Ralf,
Couturier Cyril
Publication year - 2009
Publication title -
biotechnology journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.144
H-Index - 84
eISSN - 1860-7314
pISSN - 1860-6768
DOI - 10.1002/biot.200900016
Subject(s) - bioluminescence , acceptor , förster resonance energy transfer , fluorescence , energy transfer , receptor , chemistry , luciferase , intramolecular force , ligand (biochemistry) , biophysics , green fluorescent protein , microbiology and biotechnology , biochemistry , biology , transfection , stereochemistry , gene , physics , quantum mechanics , molecular physics , condensed matter physics
Abstract We report highly sensitive bioluminescence resonance energy transfer (BRET) assays with optimized donor/acceptor couples. We combined the energy donors Renilla luciferase (Rluc) and the Rluc8 variant with the energy acceptors yellow fluorescent protein, the YPet variant and the Renilla green fluorescent protein (RGFP). Different donor/acceptor couples were tested in well‐established assays measuring ligand‐induced β‐arrestin (βARR) intramolecular rearrangements and recruitment to G protein‐coupled receptors. We show increased sensitivity with Rluc8/YPet and Rluc8/RGFP couples and measured previously undetectable BRET signals. These tools improve existing βARR assays and offer new options for the development of future BRET assays.