Agonist and antagonist TRUPATH assays for G protein-coupled receptors

Jeffrey F DiBerto; Katie Smart; Reid H J Olsen; Bryan L Roth

doi:10.1016/j.xpro.2022.101259

Agonist and antagonist TRUPATH assays for G protein-coupled receptors

STAR Protoc. 2022 Apr 1;3(2):101259. doi: 10.1016/j.xpro.2022.101259. eCollection 2022 Jun 17.

Authors

Jeffrey F DiBerto¹, Katie Smart², Reid H J Olsen², Bryan L Roth¹

Affiliations

¹ Department of Pharmacology, University of North Carolina at Chapel Hill School of Medicine, Chapel Hill, NC 27599-7365, USA.
² GPCR Pharmacology, Department of Discovery Biology, Exscientia, Oxford OX44GE, UK.

Abstract

TRUPATH is a bioluminescence resonance energy transfer-based platform for quantifying G protein-coupled receptor activity via dissociation of heterotrimeric G protein biosensors. Here, we present protocols for agonist and antagonist TRUPATH assays in the 384-well plate format, thereby providing an opportunity for higher throughput. We also provide both data analysis and quality control analyses for these assays, along with considerations for assay optimization and solutions for troubleshooting needs that may be encountered. For complete details on the use and execution of this protocol, please refer to Olsen et al. (2020).

Keywords: Biotechnology and bioengineering; Cell Biology; Cell-based Assays; High Throughput Screening; Molecular Biology; Molecular/Chemical Probes; Signal Transduction.

Publication types

Research Support, N.I.H., Extramural
Research Support, Non-U.S. Gov't

MeSH terms

Biological Assay
Biosensing Techniques* / methods
Receptors, G-Protein-Coupled* / metabolism
Signal Transduction

Substances

Receptors, G-Protein-Coupled

Abstract

Publication types

MeSH terms

Substances

Grants and funding