Regulation of RGS-RhoGEFs by Galpha12 and Galpha13 proteins.
Three mammalian Rho guanine nucleotide exchange factors (RhoGEFs), leukemia-associated RhoGEF (LARG), p115RhoGEF, and PDZ-RhoGEF, contain regulator of G-protein signaling (RGS) domains within their amino-terminal regions. These RhoGEFs link signals from heterotrimeric G12/13 protein-coupled receptors to Rho GTPase activation, leading to various cellular responses, such as actin reorganization and gene expression. The activity of these RhoGEFs is regulated by Galpha12/13 through their RGS domains. Because RhoGEFs stimulate guanine nucleotide exchange by Rho GTPases, RhoGEF activation can be measured by monitoring GTP binding to or GDP dissociation from Rho GTPases. This article describes methods used to perform reconstitution assays to measure the activity of RhoGEFs regulated by Galpha12/13.
Tanabe
Shihori
S
Department of Pharmacology, University of Illinois at Chicago, 60612, USA.
Kreutz
Barry
B
Suzuki
Nobuchika
N
Kozasa
Tohru
T
D000818:Animals; D002460:Cell Line; D044365:GTP-Binding Protein alpha Subunits, G12-G13 / Q000302:isolation & purification / Q000378:metabolism*; D020662:Guanine Nucleotide Exchange Factors / Q000302:isolation & purification / Q000378:metabolism*; D016244:Guanosine 5'-O-(3-Thiotriphosphate) / Q000378:metabolism; D006153:Guanosine Diphosphate / Q000378:metabolism; D010766:Phosphorylation; D020710:RGS Proteins / Q000378:metabolism*; D064067:Rho Guanine Nucleotide Exchange Factors; D020742:rhoA GTP-Binding Protein / Q000302:isolation & purification / Q000378:metabolism
D020662:Guanine Nucleotide Exchange Factors; D020710:RGS Proteins; D064067:Rho Guanine Nucleotide Exchange Factors; D006153:Guanosine Diphosphate; D016244:Guanosine 5'-O-(3-Thiotriphosphate); D044365:GTP-Binding Protein alpha Subunits, G12-G13; D020742:rhoA GTP-Binding Protein