title: Approaches to Characterize and Quantify Oligomerization of GPCRs.
abstract: Fluorescence resonance energy transfer (FRET) is an approach widely used to detect protein-protein interactions in live cells. This approach is based on the sensitization of an "acceptor" molecule by the energy transfer from a "donor" when there is an overlap between the emission spectrum of the "donor" and the excitation spectrum of the "acceptor" and close proximity between the two fluorophore species (in the region of 8 nm). Various methods exist to quantify FRET signals: here, we describe the application of homogeneous time-resolved FRET (htrFRET) combined with Tag-lite™ technology and its application to determine not only protein-protein interactions but also the capability of GPCR mutant variants to form homomers compared to the wild type GPCR within the plasma membrane of transfected cells.
authors.affiliation: Molecular Pharmacology Group, Institute of Molecular, Cell and Systems Biology, College of Medical, Veterinary and Life Sciences, University of Glasgow, University Avenue, Glasgow, G12 8QQ, Scotland, UK.
authors.forename: Sara
authors.initials: S
authors.lastname: Marsango
authors.forename: María José
authors.initials: MJ
authors.lastname: Varela
authors.forename: Graeme
authors.initials: G
authors.lastname: Milligan
mesh_terms: D002462:Cell Membrane / Q000378:metabolism; D031541:Fluorescence Resonance Energy Transfer / Q000379:methods*; D057809:HEK293 Cells; D006801:Humans; D016296:Mutagenesis; D009154:Mutation; D010957:Plasmids / Q000235:genetics; D055503:Protein Multimerization; D020836:Protein Structure, Quaternary; D043562:Receptors, G-Protein-Coupled / Q000737:chemistry* / Q000235:genetics
chemical_list: D043562:Receptors, G-Protein-Coupled