Monitoring G protein activation in cells with BRET

Ikuo Masuho, Kirill A. Martemyanov, Nevin A Lambert

Research output: Contribution to journalArticle

16 Citations (Scopus)

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 languageEnglish (US)
Pages (from-to)107-113
Number of pages7
JournalMethods in Molecular Biology
Volume1335
DOIs
StatePublished - Jan 1 2015
Externally publishedYes

Fingerprint

GTP-Binding Proteins
Fluorescence Resonance Energy Transfer
Protein Conformation
Second Messenger Systems
Luminescence
Proteins
Fluorescence
Pharmacology
Equipment and Supplies

Keywords

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

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics

Cite this

Monitoring G protein activation in cells with BRET. / Masuho, Ikuo; Martemyanov, Kirill A.; Lambert, Nevin A.

In: Methods in Molecular Biology, Vol. 1335, 01.01.2015, p. 107-113.

Research output: Contribution to journalArticle

Masuho, Ikuo ; Martemyanov, Kirill A. ; Lambert, Nevin A. / Monitoring G protein activation in cells with BRET. In: Methods in Molecular Biology. 2015 ; Vol. 1335. pp. 107-113.
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