Monitoring G protein activation in cells with BRET

Ikuo Masuho; Kirill A. Martemyanov; Nevin A. Lambert

doi:10.1007/978-1-4939-2914-6_8

Monitoring G protein activation in cells with BRET

Ikuo Masuho, Kirill A. Martemyanov, Nevin A. Lambert

Pharmacology and Toxicology

Research output: Contribution to journal › Article › peer-review

57 Scopus citations

Abstract

Live-cell assays based on fluorescence and luminescence are now indispensable tools for the study of G protein signaling. Assays based on fluorescence and bioluminescence resonance energy transfer (FRET and BRET) have been particularly valuable for monitoring changes in second messengers, protein-protein interactions, and protein conformation. Here, we describe a BRET assay that monitors the release of free Gβγ dimers after activation of heterotrimers containing Gα subunits from all four G protein subfamilies. This assay provides useful kinetic and pharmacological information with reasonably high throughput using a standard laboratory equipment.

Original language	English (US)
Pages (from-to)	107-113
Number of pages	7
Journal	Methods in Molecular Biology
Volume	1335
DOIs	https://doi.org/10.1007/978-1-4939-2914-6_8
State	Published - 2015

Keywords

Bioluminescence resonance energy transfer (BRET)
G protein-coupled receptors (GPCRs)
Luciferase
Transfection
Venus fluorescent protein

ASJC Scopus subject areas

Molecular Biology
Genetics

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10.1007/978-1-4939-2914-6_8

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title = "Monitoring G protein activation in cells with BRET",

abstract = "Live-cell assays based on fluorescence and luminescence are now indispensable tools for the study of G protein signaling. Assays based on fluorescence and bioluminescence resonance energy transfer (FRET and BRET) have been particularly valuable for monitoring changes in second messengers, protein-protein interactions, and protein conformation. Here, we describe a BRET assay that monitors the release of free Gβγ dimers after activation of heterotrimers containing Gα subunits from all four G protein subfamilies. This assay provides useful kinetic and pharmacological information with reasonably high throughput using a standard laboratory equipment.",

keywords = "Bioluminescence resonance energy transfer (BRET), G protein-coupled receptors (GPCRs), Luciferase, Transfection, Venus fluorescent protein",

author = "Ikuo Masuho and Martemyanov, {Kirill A.} and Lambert, {Nevin A.}",

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doi = "10.1007/978-1-4939-2914-6_8",

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T1 - Monitoring G protein activation in cells with BRET

AU - Masuho, Ikuo

AU - Martemyanov, Kirill A.

AU - Lambert, Nevin A.

N1 - Publisher Copyright: © Springer Science+Business Media New York 2015.

PY - 2015

Y1 - 2015

N2 - Live-cell assays based on fluorescence and luminescence are now indispensable tools for the study of G protein signaling. Assays based on fluorescence and bioluminescence resonance energy transfer (FRET and BRET) have been particularly valuable for monitoring changes in second messengers, protein-protein interactions, and protein conformation. Here, we describe a BRET assay that monitors the release of free Gβγ dimers after activation of heterotrimers containing Gα subunits from all four G protein subfamilies. This assay provides useful kinetic and pharmacological information with reasonably high throughput using a standard laboratory equipment.

AB - Live-cell assays based on fluorescence and luminescence are now indispensable tools for the study of G protein signaling. Assays based on fluorescence and bioluminescence resonance energy transfer (FRET and BRET) have been particularly valuable for monitoring changes in second messengers, protein-protein interactions, and protein conformation. Here, we describe a BRET assay that monitors the release of free Gβγ dimers after activation of heterotrimers containing Gα subunits from all four G protein subfamilies. This assay provides useful kinetic and pharmacological information with reasonably high throughput using a standard laboratory equipment.

KW - Bioluminescence resonance energy transfer (BRET)

KW - G protein-coupled receptors (GPCRs)

KW - Luciferase

KW - Transfection

KW - Venus fluorescent protein

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SN - 1064-3745

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EP - 113

JO - Methods in Molecular Biology

JF - Methods in Molecular Biology

ER -

Monitoring G protein activation in cells with BRET

Abstract

Keywords

ASJC Scopus subject areas

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