Lighting up multiprotein complexes: lessons from GPCR oligomerization.
Spatiotemporal characterization of protein-protein interactions (PPIs) is essential in determining the molecular mechanisms of intracellular signaling processes. In this review, we discuss how new methodological strategies derived from non-invasive fluorescence- and luminescence-based approaches (FRET, BRET, BiFC and BiLC), when applied to the study of G protein-coupled receptor (GPCR) oligomerization, can be used to detect specific PPIs in live cells. These technologies alone or in concert with complementary methods (SRET, BRET or BiFC, and SNAP-tag or TR-FRET) can be extremely powerful approaches for PPI visualization, even between more than two proteins. Here we provide a comprehensive update on all the biotechnological aspects, including the strengths and weaknesses, of new fluorescence- and luminescence-based methodologies, with a specific focus on their application for studying PPIs.
Ciruela
Francisco
F
Unitat de Farmacologia, Departament de Patologia i Terapèutica Experimental, Facultat de Medicina, IDIBELL-Universitat de Barcelona, 08907 Barcelona, Spain. fciruela@ub.edu
Vilardaga
Jean-Pierre
JP
Fernández-Dueñas
Víctor
V
D000818:Animals; D003584:Cytological Techniques; D031541:Fluorescence Resonance Energy Transfer; D006801:Humans; D008163:Luminescent Measurements / Q000379:methods*; D046912:Multiprotein Complexes / Q000737:chemistry*; D011485:Protein Binding; D025941:Protein Interaction Mapping / Q000379:methods*; D043562:Receptors, G-Protein-Coupled / Q000737:chemistry*
D046912:Multiprotein Complexes; D043562:Receptors, G-Protein-Coupled