Abstract
Intracellular signal transduced by G protein-coupled receptors (GPCRs) is tightly controlled by a guanine nucleotide-binding complex made of G protein Gα, Gβ, and Gγ subunits, as well as a growing array of regulatory and accessory proteins such as arrestins. G protein-independent β-arrestin recruitment at GPCRs is universally accepted as the canonical interactor system and it has been found to be a powerful tracker of most GPCRs activation. Pharmacological concepts have evolved remarkably after the finding that different ligands, binding at the same receptor, can selectively activate specific subsets of signaling pathways among all pathways activated by balanced ligands. This new paradigm referred to as functional selectivity or biased signaling, has opened new avenues for the design of tailored drugs with enhanced therapeutic efficacies and reduced side effects. Here, we describe a unique platform for the interrogation of GPCR using a transcriptional-based assay to measure transient β-arrestin recruitment called Tango.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Luttrell LM, Gesty-Palmer D (2010) Beyond desensitization: physiological relevance of arrestin-dependent signaling. Pharmacol Rev 62(2):305–330. https://doi.org/10.1124/pr.109.002436
Katritch V, Cherezov V, Stevens RC (2013) Structure-function of the G protein-coupled receptor superfamily. Annu Rev Pharmacol Toxicol 53:531–556. https://doi.org/10.1146/annurev-pharmtox-032112-135923
Urban JD, Clarke WP, von Zastrow M, Nichols DE, Kobilka B, Weinstein H, Javitch JA, Roth BL, Christopoulos A, Sexton PM, Miller KJ, Spedding M, Mailman RB (2007) Functional selectivity and classical concepts of quantitative pharmacology. J Pharmacol Exp Ther 320(1):1–13. https://doi.org/10.1124/jpet.106.104463
Kroeze WK, Sassano MF, Huang XP, Lansu K, McCorvy JD, Giguere PM, Sciaky N, Roth BL (2015) PRESTO-Tango as an open-source resource for interrogation of the druggable human GPCRome. Nat Struct Mol Biol. https://doi.org/10.1038/nsmb.3014
Barnea G, Strapps W, Herrada G, Berman Y, Ong J, Kloss B, Axel R, Lee KJ (2008) The genetic design of signaling cascades to record receptor activation. Proc Natl Acad Sci U S A 105(1):64–69. https://doi.org/10.1073/pnas.0710487105
Jordan M, Schallhorn A, Wurm FM (1996) Transfecting mammalian cells: optimization of critical parameters affecting calcium-phosphate precipitate formation. Nucleic Acids Res 24(4):596–601
Baker JM, Boyce FM (2014) High-throughput functional screening using a homemade dual-glow luciferase assay. J Vis Exp (88). https://doi.org/10.3791/50282
Acknowledgments
This work was supported by the Canadian Institutes of Health Research MOP142219.
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2019 Springer Science+Business Media, LLC, part of Springer Nature
About this protocol
Cite this protocol
Laroche, G., Giguère, P.M. (2019). Measurement of β-Arrestin Recruitment at GPCRs Using the Tango Assay. In: Tiberi, M. (eds) G Protein-Coupled Receptor Signaling. Methods in Molecular Biology, vol 1947. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-9121-1_14
Download citation
DOI: https://doi.org/10.1007/978-1-4939-9121-1_14
Published:
Publisher Name: Humana Press, New York, NY
Print ISBN: 978-1-4939-9120-4
Online ISBN: 978-1-4939-9121-1
eBook Packages: Springer Protocols