Detection of GPCR/beta-Arrestin Interactions in Live Cells Using Bioluminescence Resonance Energy Transfer Technology

Abstract

Bioluminescence resonance energy transfer (BRET) is a powerful and increasingly popular technique for studying protein-protein interactions in live cells and real time. In particular, there has been considerable interest in the ability to monitor interactions between G protein-coupled receptors (GPCRs) and proteins that serve as key regulators of receptor function, such as beta-arrestin. The BRET methodology involves heterologous co-expression of genetically fused proteins that link one protein of interest (e.g., a GPCR) to a bioluminescent donor enzyme and a second protein of interest (e.g., beta-arrestin) to an acceptor fluorophore. If the fusion proteins are in close proximity, resonance ..