title: Determining Rho GTPase activity by an affinity-precipitation assay.
abstract: The Rho GTPases are members of the Ras superfamily of GTPases that are pivotal regulators of the actin cytoskeleton. They also contribute to other cellular processes such as gene transcription, cell polarity, microtubule dynamics, cell cycle progression and vesicle trafficking. Most Rho GTPases act as molecular switches cycling between an "active" GTP-bound form and an "inactive" GDP-bound form. Hence, to elucidate the mechanisms by which Rho GTPases regulate cellular responses, an important parameter to determine is the GTP-loading of each Rho family member in cells under different conditions. Here we describe a biochemical technique to assess this based on affinity-precipitation of the GTP-bound form from whole cell lysates.
authors.affiliation: Randall Division of Cell and Molecular Biophysics, King's College London, London, UK.
authors.forename: Narendra
authors.initials: N
authors.lastname: Suryavanshi
authors.forename: Anne J
authors.initials: AJ
authors.lastname: Ridley
mesh_terms: D008841:Actin Cytoskeleton / Q000378:metabolism*; D002453:Cell Cycle / Q000235:genetics; D006160:Guanosine Triphosphate / Q000378:metabolism; D006801:Humans; D008967:Molecular Biology / Q000379:methods*; D014158:Transcription, Genetic; D020741:rho GTP-Binding Proteins / Q000302:isolation & purification* / Q000378:metabolism
chemical_list: D006160:Guanosine Triphosphate; D020741:rho GTP-Binding Proteins