title: Tango assay for ligand-induced GPCR-β-arrestin2 interaction: Application in drug discovery.
abstract: G protein-coupled receptors (GPCRs) are widely known to modulate almost all physiological functions and have been demonstrated over the time as therapeutic targets for wide gamut of diseases. The design and implementation of high-throughput GPCR-based assays that permit the efficient screening of large compound libraries to discover novel drug candidates are essential for a successful drug discovery endeavor. Usually, GPCR-based functional assays depend primarily on the measurement of G protein-mediated second messenger generation. However, with advent of advanced molecular biology tools and increased understanding of GPCR signal transduction, many G protein-independent pathways such as β-arrestin translocation are being utilized to detect the activity of GPCRs. These assays provide additional information on functional selectivity (also known as biased agonism) of compounds that could be harnessed to develop pathway-selective drug candidates to reduce the adverse effects associated with given GPCR target. In this chapter, we describe the basic principle, detailed methodologies and assay setup, result analysis and data interpretations of the β-arrestin2 Tango assay, and its comparison with cell-based G protein-dependent GPCR assays, which could be employed in a simple academic setup to facilitate GPCR-based drug discovery.
authors.affiliation: Division of Pharmacology, CSIR-Central Drug Research Institute, Lucknow, Uttar Pradesh, India.
authors.forename: Shalini
authors.initials: S
authors.lastname: Dogra
authors.affiliation: Division of Pharmacology, CSIR-Central Drug Research Institute, Lucknow, Uttar Pradesh, India.
authors.forename: Chandan
authors.initials: C
authors.lastname: Sona
authors.affiliation: Division of Pharmacology, CSIR-Central Drug Research Institute, Lucknow, Uttar Pradesh, India.
authors.forename: Ajeet
authors.initials: A
authors.lastname: Kumar
authors.affiliation: Division of Pharmacology, CSIR-Central Drug Research Institute, Lucknow, Uttar Pradesh, India.
authors.forename: Prem N
authors.initials: PN
authors.lastname: Yadav
mesh_terms: D004305:Dose-Response Relationship, Drug; D004353:Drug Evaluation, Preclinical / Q000379:methods; D057809:HEK293 Cells; D006801:Humans; D020128:Inhibitory Concentration 50; D008024:Ligands; D043562:Receptors, G-Protein-Coupled / Q000819:agonists / Q000037:antagonists & inhibitors / Q000378:metabolism*; D015398:Signal Transduction / Q000187:drug effects; D000071560:beta-Arrestin 2 / Q000378:metabolism*
chemical_list: D008024:Ligands; D043562:Receptors, G-Protein-Coupled; D000071560:beta-Arrestin 2
keywords: Drug discovery; Functional selectivity; G protein-coupled receptors (GPCRs); High-throughput screening; Tango assay; β-arrestin