GPCR oligomerization analysis by means of BRET and dFRAP

Abstract

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.