Protocol to Study β-Arrestin Recruitment by CB1 and CB2 Cannabinoid Receptors

Marjolein Soethoudt, Noortje van Gils, Mario van der Stelt, Laura H Heitman

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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.

Original languageEnglish
Pages (from-to)103-11
Number of pages9
JournalMethods in Molecular Biology
Publication statusPublished - 2016

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