Detection of receptor heteromers involving dopamine receptors by the sequential BRET-FRET technology

Gemma Navarro, Peter J. McCormick, Josefa Mallol, Carme Lluís, Rafael Franco, Antoni Cortés, Vicent Casadó, Enric I. Canela, Sergi Ferré

Research output: Contribution to journalArticlepeer-review

8 Citations (Scopus)


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.
Original languageEnglish
Pages (from-to)95-105
Number of pages11
JournalMethods in Molecular Biology
Publication statusPublished - 1 Jan 2013

Cite this