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Determining the Activation of Rho as an Index of Receptor Coupling to G12/13 Proteins

  • Michio Nakaya
  • Mina Ohba
  • Motohiro Nishida
  • Hitoshi KuroseEmail author
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 746)

Abstract

Heterotrimeric G proteins are composed of α, β, and γ subunits. G proteins can be activated by a large number of cell-surface hepathelical receptors and can transduce signals from these receptors to various intracellular signaling molecules. When G protein-coupled receptors are bound by their cognate ligand, interaction with specific subtypes of G protein leads to dissociation of the α subunit of the heterotrimeric G protein from the βγ dimer, and both Gα-GTP and Gβγ are capable of initiating their own signal transduction pathways. G proteins are functionally divided into four groups based on the nature of α subunit into Gs, Gi, Gq, and G12 families. The members of the G12 subfamily are G12 and G13. Increasing evidence indicates that G12/13 proteins play critical roles in various physiological functions. G12 and G13 regulate the small GTPase Rho through modulation of guanine nucleotide exchange factor (RhoGEF) activity to regulate various cellular responses, such as cytoskeletal changes and cell growth. Therefore, Rho activity can often represent a sensitive marker of G12/13 activity. Here, we describe the Rho activation assay to monitor the activity of G12/13 proteins.

Key words

G12 Rho Pull-down Cardiomyocyte 

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Copyright information

© Springer Science+Business Media, LLC 2011

Authors and Affiliations

  • Michio Nakaya
  • Mina Ohba
  • Motohiro Nishida
  • Hitoshi Kurose
    • 1
    Email author
  1. 1.Department of Pharmacology and Toxicology, Graduate School of Pharmaceutical SciencesKyushu UniversityFukuokaJapan

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