title: Improved donor/acceptor BRET couples for monitoring beta-arrestin recruitment to G protein-coupled receptors.
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 beta-arrestin (betaARR) 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 betaARR assays and offer new options for the development of future BRET assays.
authors: Institut Cochin, Université Paris Descartes, CNRS (UMR 8104), Paris, France.
authors: Maud
authors: M
authors: Kamal
authors: Marcel
authors: M
authors: Marquez
authors: Virginie
authors: V
authors: Vauthier
authors: Audrey
authors: A
authors: Leloire
authors: Philippe
authors: P
authors: Froguel
authors: Ralf
authors: R
authors: Jockers
authors: Cyril
authors: C
authors: Couturier
mesh_terms: D019390:Arrestins / Q000032:analysis* / Q000737:chemistry*; D031541:Fluorescence Resonance Energy Transfer / Q000379:methods*; D008163:Luminescent Measurements / Q000379:methods*; D011485:Protein Binding; D025941:Protein Interaction Mapping / Q000379:methods*; D043562:Receptors, G-Protein-Coupled / Q000032:analysis* / Q000737:chemistry*; D000071557:beta-Arrestins
chemical_list: D019390:Arrestins; D043562:Receptors, G-Protein-Coupled; D000071557:beta-Arrestins