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

Xiaoyuan Ma; Rob Leurs; Henry F. Vischer

doi:10.1007/978-1-0716-1221-7_16

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 proceeding › Chapter › Academic › peer-review

Abstract

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 language	English
Title of host publication	G Protein-Coupled Receptor Screening Assays
Subtitle of host publication	Methods and Protocols
Editors	Sofia Aires M. Martins, Duarte Miguel F. Prazeres
Publisher	Humana Press Inc
Pages	233-248
Number of pages	16
Edition	2nd
ISBN (Electronic)	9781071612217
ISBN (Print)	9781071612200, 9781071612231
DOIs	https://doi.org/10.1007/978-1-0716-1221-7_16
Publication status	Published - 2021

Publication series

Name	Methods in Molecular Biology
Publisher	Springer
Volume	2268
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:
Copyright 2021 Elsevier B.V., All rights reserved.

Funding

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).

Funders	Funder number
CSC Chinese	201703250074
Promega
Korea University

Keywords

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

UN SDGs

This output contributes to the following UN Sustainable Development Goals (SDGs)

Access to Document

10.1007/978-1-0716-1221-7_16

Persistent URL (handle)

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Cite this

Ma, X., Leurs, R., & Vischer, H. F. (2021). NanoLuc-Based Methods to Measure β-Arrestin2 Recruitment to G Protein-Coupled Receptors. In S. A. M. Martins, & D. M. F. Prazeres (Eds.), G Protein-Coupled Receptor Screening Assays: Methods and Protocols (2nd ed., pp. 233-248). (Methods in Molecular Biology; Vol. 2268). Humana Press Inc. https://doi.org/10.1007/978-1-0716-1221-7_16

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NanoLuc-Based Methods to Measure β-Arrestin2 Recruitment to G Protein-Coupled Receptors. / Ma, Xiaoyuan; Leurs, Rob ; Vischer, Henry F.
G Protein-Coupled Receptor Screening Assays: Methods and Protocols. ed. / Sofia Aires M. Martins; Duarte Miguel F. Prazeres. 2nd. ed. Humana Press Inc, 2021. p. 233-248 (Methods in Molecular Biology; Vol. 2268).

Research output: Chapter in Book / Report / Conference proceeding › Chapter › Academic › peer-review

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

Abstract

Publication series

Bibliographical note

Funding

Keywords

UN SDGs

Access to Document

Persistent URL (handle)

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Cite this