Abstract
Cannabinoid CB1 and CB2 receptors are G-protein-coupled receptors (GPCRs) that recruit β-arrestins upon activation by (partial) agonists. β-Arrestin recruitment is induced by phosphorylation of their C-terminal tails, and is associated with the termination of GPCR signaling; yet, it may also activate cellular signaling pathways independent of G-proteins. Here, we describe a detailed protocol to characterize the potency and efficacy of ligands to induce or inhibit β-arrestin recruitment to the human CB1 and CB2 receptors, by using the PathHunter® assay. The latter is a cellular assay that can be performed in plates with 384-wells. The PathHunter® assay makes use of β-galactosidase complementation, and has a chemiluminescent readout. We used this assay to characterize a set of reference ligands (both agonists and antagonists) on human CB1 and CB2 receptors.
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The authors wish to thank DiscoveRx for their technical assistance.
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Soethoudt, M., van Gils, N., van der Stelt, M., Heitman, L.H. (2016). Protocol to Study β-Arrestin Recruitment by CB1 and CB2 Cannabinoid Receptors. In: Maccarrone, M. (eds) Endocannabinoid Signaling. Methods in Molecular Biology, vol 1412. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-3539-0_11
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DOI: https://doi.org/10.1007/978-1-4939-3539-0_11
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Publisher Name: Humana Press, New York, NY
Print ISBN: 978-1-4939-3537-6
Online ISBN: 978-1-4939-3539-0
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