Application of BRET for Studying G Protein-Coupled Receptors
Agnieszka A. Kaczor, Magdalena Makarska-Bialokoz, Jana Selent, Rocio A. de la Fuente, Maria Marti-Solano and Marian Castro
Affiliation: Department of Synthesis and Chemical Technology of Pharmaceutical Substances with Computer Modeling Lab, Faculty of Pharmacy with Division of Medical Analytics, Medical University of Lublin, 4A Chodzki St., PL-20093 Lublin, Poland.
Keywords: BRET, G protein-coupled receptors, G protein-coupled receptor dimers.
G protein-coupled receptors (GPCRs) constitute one of the largest classes of cell surface receptors. GPCR
biology has been a subject of widespread interest owing to the functional relevance of these receptors and their potential
importance in the development of new drugs. At present, over 30% of all launched drugs target these receptors. GPCRs
have been considered for a long time to function as monomeric entities and the idea of GPCR dimerization and
oligomerization was initially accepted with disbelief. However, a significant amount of experimental and molecular
modeling evidence accumulated during the last several years suggests that the process of GPCRs dimer or oligomer
formation is a general phenomenon, in some cases even essential for receptor function. Among the many methods to study
GPCR dimerization and oligomerization, modern biophysical techniques such as those based on resonance energy transfer
(RET) and particularly bioluminescence resonance energy transfer (BRET) have played a leading role. RET methods are
commonly applied as non-destructive indicators of specific protein-protein interactions (PPIs) in living cells. Data from
numerous BRET experiments support the idea that the process of GPCR oligomerization may be relevant in many
physiological and pathological conditions. The application of BRET to the study of GPCRs is not only limited to the
assessment of receptor oligomerization but also expands to the investigation of the interactions of GPCRs with other
proteins, including G proteins, G protein-coupled receptor kinases, β-arrestins or receptor tyrosine kinases, as well as to
the characterization of GPCR activation and signaling. In this review, we briefly summarize the fundaments of BRET,
discuss new trends in this technology and describe the wide range of applications of BRET to study GPCRs.
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