title: 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.
authors.affiliation: Unitat de Farmacologia, Departament Patologia i Terapèutica Experimental, Facultat de Medicina, IDIBELL, L'Hospitalet de Llobregat, Universitat de Barcelona, Barcelona, 08907, Spain.
authors.forename: Francisco
authors.initials: F
authors.lastname: Ciruela
authors.forename: Víctor
authors.initials: V
authors.lastname: Fernández-Dueñas
mesh_terms: D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism*; D060807:Bioluminescence Resonance Energy Transfer Techniques; D036681:Fluorescence Recovery After Photobleaching; D015870:Gene Expression; D017930:Genes, Reporter; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism*; D057809:HEK293 Cells; D006801:Humans; D008156:Luciferases / Q000235:genetics / Q000378:metabolism*; D049408:Luminescent Agents / Q000737:chemistry; D008164:Luminescent Proteins / Q000235:genetics / Q000378:metabolism*; D010957:Plasmids / Q000737:chemistry / Q000378:metabolism; D011485:Protein Binding; D054730:Protein Interaction Domains and Motifs; D055503:Protein Multimerization; D021381:Protein Transport; D043705:Receptor, Adenosine A2A / Q000235:genetics / Q000378:metabolism*; D017448:Receptors, Dopamine D2 / Q000235:genetics / Q000378:metabolism*; D014162:Transfection
chemical_list: D001426:Bacterial Proteins; D049408:Luminescent Agents; D008164:Luminescent Proteins; D043705:Receptor, Adenosine A2A; D017448:Receptors, Dopamine D2; C054562:yellow fluorescent protein, Bacteria; D049452:Green Fluorescent Proteins; D008156:Luciferases