Demonstration of Improvements to the Bioluminescence Resonance Energy Transfer (BRET) Technology for the Monitoring of G Protein-Coupled Receptors in Live Cells
Martina Kocan, Heng B See, Ruth M Seeber, Karin A Eidne, Kevin DG Pfleger
Journal of Biomolecular Screening | SAGE PUBLICATIONS INC | Published : 2008
The bioluminescence resonance energy transfer (BRET) technique has become extremely popular for studying protein-protein interactions in living cells and real time. Of particular interest is the ability to monitor interactions between G protein-coupled receptors, such as the thyrotropin-releasing hormone receptor (TRHR), and proteins critical for regulating their function, such as beta-arrestin. Using TRHR/beta-arrestin interactions, we have demonstrated improvements to all 3 generations of BRET (BRET(1), BRET(2), and eBRET) by using the novel forms of luciferase, Rluc2 and Rluc8, developed by the Gambhir laboratory. Furthermore, for the 1st time it was possible to use the BRET2 system to de..View full abstract
Awarded by National Health and Medical Research Council (NHMRC) of Australia
Awarded by NHMRC Peter Doherty Fellowship
We thank Yuann S. Heng for expert technical assistance and Matthew B. Dalrymple for critical reading of the article. We are grateful to Andreas Loening and Sanjiv Gambhir (Stanford University. Stanford, CA), Atsushi Miyawaki (RIKEN Brain Science Institute. Wako-city, Japan), and Michel Bouvier (Department of Biochemistry, Universite de Montreal, Montreal, Quebec, Canada) for providing cDNA constructs. This work was funded by the National Health and Medical Research Council (NHMRC) of Australia (Project Grant #404087). Kevin D. G. Pfleger is supported by an NHMRC Peter Doherty Fellowship (#353709).