Improved donor/acceptor BRET couples for monitoring ß-arrestin recruitment to G protein-coupled receptors

Maud Kamal (1) , Marcel Marquez (2) , Virginie Vauthier (1) , Audrey Leloire (2) , Philippe Froguel (2) , Ralf Jockers (1) , Cyril Couturier (2)

1 UMR_S567 / UMR 8104 - Institut Cochin
2 GMM - Génétique des maladies multifactorielles

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 (YFP), the YPet variant and the Renilla green fluorescent protein (RGFP). Different donor/acceptor couples were tested in well-established assays measuring ligand-induced ß-arrestin (ßARR) intramolecular rearrangements and recruitment to G protein-coupled receptors (GPCRs). We show increased sensitivity with Rluc8/YPet and Rluc8/RGFP couples and measured previously undetectable BRET signals. These tools improve existing ßARR assays and offer new options for the development of future BRET assays.

Mots clés: BRET; RGFP; YPet; GPCR; ßarrestin

Maud Kamal, Marcel Marquez, Virginie Vauthier, Audrey Leloire, Philippe Froguel, et al.. Improved donor/acceptor BRET couples for monitoring ß-arrestin recruitment to G protein-coupled receptors. Biotechnology Journal, 2009, 4 (8), pp.1337. ⟨10.1002/biot.200900016⟩. ⟨hal-00495062⟩