Applications of BRET to study dynamic G-protein coupled receptor interactions in living cells

Summary Protein-protein interactions are fundamental processes for many biological systems including those involving the superfamily of G-protein coupled receptors (GPCRs). When addressing key questions concerning the regulation of GPCR-protein complexes and their functional significance, the development and refinement of non-invasive techniques to study these interactions will be of great value. One such technique, bioluminescence resonance energy transfer (BRET), is a recently described biophysical method that represents a powerful tool with which to measure protein-protein interactions in live cells, in real time. This minireview highlights the impact that evolving techniques such as BRET have had on the study of dynamic protein interactions involving GPCRs. In particular, the application of BRET to the study of protein interactions involving the receptors for hypothalamic peptide hormones, thyrotropin-releasing hormone (TRH) and gonadotropin-releasing hormone (GnRH), will be discussed. Using these receptors, BRET has successfully been used to demonstrate formation of both agonist-dependent and independent GPCR-GPCR complexes (oligomerization) and the agonist-dependent interaction of GPCRs with their intracellular adaptor protein partners, the arrestins. In summary, BRET is a highly snnsitive method that will not only aid in advancing our understanding of GPCR signalling and trafficking bout coud also potentially lead to the development of novel therapeutics that target these GPCR-protein complexes.