GPCR oligomerization analysis by means of BRET and dFRAP

The spatiotemporal characterization of protein-protein interactions is essential to understand nearly all cellular events. Several methodological strategies derived from noninvasive luminescence- and fluorescence-based approaches allow the detection of specific protein-protein interactions in living cells. Here, we describe the application of bioluminescence resonance energy transfer (BRET) and donor fluorescent recovery after photobleaching (dFRAP) approaches to the study of G protein-coupled receptor (GPCR) oligomerization. These technologies alone - or in combination with complementary methods - can become extremely powerful approaches for visualizing these cellular protein-protein interactions, even between more than two proteins. Overall, these methods might become extremely important tools in GPCR drug discovery.