NanoLuc-Based Methods to Measure β-Arrestin2 Recruitment to G Protein-Coupled Receptors

Xiaoyuan Ma, Rob Leurs, Henry F. Vischer*

*Corresponding author for this work

Research output: Chapter in Book / Report / Conference proceedingChapterAcademicpeer-review


Cytosolic β-arrestins are key regulators of G protein-coupled receptors (GPCRs) by sterically uncoupling G protein activation, facilitating receptor internalization, and/or acting as G protein-independent signaling scaffolds. The current awareness that GPCR ligands may display bias toward G protein signaling or β-arrestin recruitment makes β-arrestin recruitment assays important additions to the drug discovery toolbox. This chapter describes two NanoLuc-based methods to monitor β-arrestin2 recruitment to the human histamine H1 receptor by measuring bioluminescence resonance energy transfer and enzyme-fragment complementation in real-time on living cells with reasonable high throughput. In addition to the detection of agonism, both assay formats can be used to qualitatively evaluate the binding kinetics of antihistamines on the human histamine H1 receptor.

Original languageEnglish
Title of host publicationG Protein-Coupled Receptor Screening Assays
Subtitle of host publicationMethods and Protocols
EditorsSofia Aires M. Martins, Duarte Miguel F. Prazeres
PublisherHumana Press Inc
Number of pages16
ISBN (Electronic)9781071612217
ISBN (Print)9781071612200, 9781071612231
Publication statusPublished - 2021

Publication series

NameMethods in Molecular Biology
ISSN (Print)1064-3745
ISSN (Electronic)1940-6029

Bibliographical note

Funding Information:
The NanoBiT starter kit plasmids were kindly provided by Promega Corporation (Madison, Wisconsin, USA). The ?-arrestin2 Nano-BiT constructs were kindly provided by Dr. J.Y. Seong (Korea University, Seoul, Republic of Korea). The Venus-?-arrestin2-pcDNA3 construct was kindly provided by Dr. V.V. Gurevich (Van-derbilt University School of Medicine, Nashville, Tennessee, USA). X.M. is supported by a CSC Chinese scholarship grant (201703250074).

Publisher Copyright:
© 2021, Springer Science+Business Media, LLC, part of Springer Nature.

Copyright 2021 Elsevier B.V., All rights reserved.


  • Bioluminescence resonance energy transfer (BRET)
  • Enzyme-fragment complementation (EFC)
  • GPCR
  • Luciferase
  • Protein-protein interaction (PPI)
  • β-Arrestin


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