Use of BRET to Measure β-Arrestin Recruitment at Oxytocin and Vasopressin Receptors.

Bioluminescence resonance energy transfer (BRET) is a cutting-edge biophysical technique used for exploring G protein-coupled receptor (GPCR) pharmacology. BRET relies on the nonradiative energy transfer from a luciferase energy donor to an acceptor fluorophore after oxidation of a luciferase substrate. This energy transfer occurs only if the donor and acceptor are within close proximity. Over the past few years, BRET has been successfully applied to study GPCR oligomerization as well as interactions of receptors with G proteins, G protein-coupled receptor kinases (GRKs), or β-arrestins. Herein, we describe how BRET can be applied to study signaling at the oxytocin receptor (OTR) and vasopressin receptors, thereby enabling the identification of (biased) ligands and molecular probes for investigating receptor functionality.