title: Quantifying sphingosine-1-phosphate-dependent activation of the RhoGTPases.
abstract: The GPCR-coupled sphingosine-1-phosphate (S1P) receptors regulate a number of important cell -functions, including proliferation, migration, and adhesion. Since these processes require dynamic regulation of the actin cytoskeleton, the ability to monitor S1P-dependent activation of the Rho family GTPases is critical for our understanding of S1P signaling. Herein, we provide methods for the GST pull-down-based assay used to measure Rho, Rac, and Cdc42 activity in cultured cells treated with S1P.
authors.affiliation: Laboratory MedicineDepartment of Pathology and Laboratory Medicine, University of North Carolina, Chapel Hill, NC, USA.
authors.forename: Matt D
authors.initials: MD
authors.lastname: Medlin
authors.forename: Joan M
authors.initials: JM
authors.lastname: Taylor
authors.forename: Christopher P
authors.initials: CP
authors.lastname: Mack
mesh_terms: D004789:Enzyme Activation; D005455:Fluorescent Antibody Technique; D008246:Lysophospholipids / Q000378:metabolism*; D013110:Sphingosine / Q000031:analogs & derivatives* / Q000378:metabolism; D020741:rho GTP-Binding Proteins / Q000378:metabolism*
chemical_list: D008246:Lysophospholipids; C060506:sphingosine 1-phosphate; D020741:rho GTP-Binding Proteins; D013110:Sphingosine