title: Detection of receptor heteromers involving dopamine receptors by the sequential BRET-FRET technology.
abstract: Until very recently, dopamine receptors, like other G-protein-coupled receptors, were believed to function as individual units on the cell surface. Now it has been described by several groups including ours that dopamine receptors not only function as homomers but also form heteromers with other receptors at the membrane level. Bioluminescence and fluorescence resonance energy transfer (BRET and FRET) based techniques have been very useful to determine the interaction between two receptors, but to demonstrate the existence of higher-order complexes involving more than two molecules requires more sophisticated techniques. Combining BRET and FRET in the Sequential BRET-FRET (SRET) technique permits heteromers formed by three different proteins to be identified. In SRET experiments, the oxidation of a Renilla Luciferase substrate triggers acceptor excitation by BRET and subsequent energy transfer to a FRET acceptor. Using this methodology here we describe the heteromerization between adenosine A(2A), dopamine D(2), and cannabinoids CB(1) receptors in living cells.
authors.affiliation: Department of Biochemistry and Molecular Biology, University of Barcelona, Barcelona, Spain.
authors.forename: Gemma
authors.initials: G
authors.lastname: Navarro
authors.forename: Peter J
authors.initials: PJ
authors.lastname: McCormick
authors.forename: Josefa
authors.initials: J
authors.lastname: Mallol
authors.forename: Carme
authors.initials: C
authors.lastname: Lluís
authors.forename: Rafael
authors.initials: R
authors.lastname: Franco
authors.forename: Antoni
authors.initials: A
authors.lastname: Cortés
authors.forename: Vicent
authors.initials: V
authors.lastname: Casadó
authors.forename: Enric I
authors.initials: EI
authors.lastname: Canela
authors.forename: Sergi
authors.initials: S
authors.lastname: Ferré
mesh_terms: D060807:Bioluminescence Resonance Energy Transfer Techniques / Q000379:methods*; D002462:Cell Membrane / Q000378:metabolism; D031541:Fluorescence Resonance Energy Transfer / Q000379:methods*; D057809:HEK293 Cells; D006801:Humans; D055503:Protein Multimerization; D020836:Protein Structure, Quaternary; D021381:Protein Transport; D011954:Receptors, Dopamine / Q000032:analysis* / Q000737:chemistry* / Q000378:metabolism; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000378:metabolism; D013997:Time Factors
chemical_list: D011954:Receptors, Dopamine; D011993:Recombinant Fusion Proteins