Abstract
The melatonin receptor subfamily belongs to the G protein-coupled receptor superfamily and consists of three members in mammals, MT1, MT2, and GPR50. These receptors can interact with each other to form homo- and heterodimers that are part of larger molecular complexes composed of G proteins, β-arrestins, and other membrane and cytosolic proteins. BRET (bioluminescence resonance energy transfer) is a versatile technique to follow protein-protein interactions on the nanometer scale, in real time, in living cells, which contributed largely to our understanding of the function of melatonin receptors. In this chapter, we describe our BRET protocols for melatonin receptors, which can also be applied to other GPCRs.
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Acknowledgments
We acknowledge the members of the Jockers lab (Institut Cochin, France), the Department of Anatomy (Kyorin University, Japan), and the Yoshimi lab (National Cancer Center Research Institute, Japan) for the kind support. This work was supported by grants from the Vehicle Racing Commemorative Foundation (to A.O.), Daiwa Securities Health Foundation (to A.O.), and from the Fondation Recherche Médicale (Equipe FRM 2019, EQU201903008055), The French National Research Agency (ANR) ANR-19-CE16-0025-01 (Mito-GPCR), ANR-21-CE18-0023-01 (alloGLP1R), Recherches Partenariales et Innovation Biomédicale 2012 “MED-HET-REC-2”, INSERM; CNRS, La Ligue Contre le Cancer N/Ref: RS19/75-127 (to R.J.).
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Oishi, A., Jockers, R. (2022). Measuring Protein-Protein Interactions of Melatonin Receptors by Bioluminescence Resonance Energy Transfer (BRET). In: Jockers, R., Cecon, E. (eds) Melatonin. Methods in Molecular Biology, vol 2550. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-2593-4_26
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DOI: https://doi.org/10.1007/978-1-0716-2593-4_26
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