Abstract
Bioluminescence resonance energy transfer (BRET) is a very sensitive technique employed to study protein–protein interactions, including G-protein-coupled receptors (GPCRs) hetero- and homo-dimerization. Recently, BRET has also been used to investigate the interaction between GPCRs (e.g., β2 adrenergic receptor, muscarinic M2 receptor, dopaminergic D2 receptor) and non-visual arrestins. Here a BRET protocol is described to investigate interactions between the kappa opioid receptor (KOR) and non visual arrestins (arrestin-2 and arrestin-3) in HEK-293 cells, both under basal conditions and after exposure to KOR ligands.
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Acknowledgements
The author would like to acknowledge Prof. V. Gurevich for providing the plasmids backbones and for the precious advice in setting up the protocol presented in this chapter; authors would like also to acknowledge Prof. C. Chavkin, Dr. Selena S. Shattauer and Mrs. Jamie R. Kuhar for their practical support with KOR sub-cloning into the donor plasmid and for their critical review of the experimental procedures presented in this chapter.
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Bedini, A. (2015). Bioluminescence Resonance Energy Transfer (BRET) to Detect the Interactions Between Kappa Opioid Receptor and Non visual Arrestins. In: Spampinato, S. (eds) Opioid Receptors. Methods in Molecular Biology, vol 1230. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-1708-2_9
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DOI: https://doi.org/10.1007/978-1-4939-1708-2_9
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