Measurement of GTP-binding and GTPase activity of heterotrimeric Gα proteins

Swarup Roy Choudhury, Corey S. Westfall, Dieter Hackenberg, Sona Pandey

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

21 Scopus citations

Abstract

Heterotrimeric G-proteins are important signaling intermediates in all eukaryotes. These proteins link signal perception by a cell surface localized receptor to the downstream effectors of a given signaling pathways. The minimal core of the heterotrimeric G-protein complex consists of Gα, Gβ, and Gγ subunits, the G protein coupled receptor (GPCR) and the regulator of G-protein signaling (RGS) proteins. Signal transduction by heterotrimeric G-proteins is controlled by the distinct biochemical activities of Gα protein, which binds and hydrolyses GTP. Evaluation of the rate of GTP binding, the rate of GTP hydrolysis, and the rate of GTP/GDP exchange on Gα protein are required to better understand the mechanistic aspects of heterotrimeric G-protein signaling, which remains significantly limited for the plant G-proteins. Here we describe the optimized methods for measurement of the distinct biochemical activities of the Arabidopsis Gα protein.

Original languageEnglish
Title of host publicationG Protein-Coupled Receptor Signaling in Plants
Subtitle of host publicationMethods and Protocols
PublisherHumana Press Inc.
Pages13-20
Number of pages8
ISBN (Print)9781627035316
DOIs
StatePublished - 2013

Publication series

NameMethods in Molecular Biology
Volume1043
ISSN (Print)1064-3745

Keywords

  • Arabidopsis GPA1
  • GDP/GTP exchange
  • GTP hydrolysis
  • GTP-binding
  • GTPase activity
  • Gα protein
  • Heterotrimeric G-protein

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