Related Experiment Video
Updated: Aug 18, 2026

RhoC GTPase Activation Assay
Published on: August 22, 2010
Muscarinic receptor-stimulated cytosol-membrane translocation of RhoA
Abstract:
Receptor-mediated phospholipase D activation in human embryonic kidney (HEK) cells stably expressing the m3 muscarinic acetylcholine receptor (mAChR) apparently involves the small G protein RhoA. Here, activation of RhoA was examined by measuring cytosol-membrane translocation, which is a sign of RhoA activation. RhoA translocation was induced by guanosine 5'-O-(3-thio)triphosphate in digitonin-permeabilized HEK cells, and in intact cells by the agonist-activated mAChR and by direct activation of heterotrimeric G proteins. RhoA translocation was also induced by the phosphotyrosine phosphatase inhibitor pervanadate, while the tyrosine kinase inhibitors tyrphostin 23 and genistein inhibited the mAChR-induced RhoA translocation. These data suggest that translocation and thus activation of RhoA by the G protein-coupled m3 mAChR in HEK cells apparently involves a tyrosine kinase-dependent reaction.
More Related Videos
13:51Detection of Small GTPase Prenylation and GTP Binding Using Membrane Fractionation and GTPase-linked Immunosorbent Assay
Published on: November 11, 2018
12:35Optogenetic Inhibition of Rho1-Mediated Actomyosin Contractility Coupled with Measurement of Epithelial Tension in Drosophila Embryos
Published on: April 14, 2023
Related Concept Videos
Rab Proteins
Rab proteins switch between a cytosolic, GDP-bound inactive state and a membrane-anchored, GTP-bound active state. By themselves, Rabs show slow rates of GDP/GTP exchange and GTP hydrolysis. Thus, Rab proteins are considered...
The Ras Gene
Ras is a superfamily...
Cell Motility through Blebbing
Blebbing Through the Matrix
In multicellular...
Cell Polarization by Rho Proteins
Small GTPases - Ras and Rho
Three regulatory proteins control their activity:
MAPK Signaling Cascades