Purification and functional analysis of Ric-8A: a guanine nucleotide exchange factor for G-protein alpha subunits.
Ric-8A (synembryn) has been shown to accelerate the in vitro guanine nucleotide exchange activities of most G-protein alpha subunits (with the exception of Galphas). Methods are presented in this article for the purification of Ric-8A and functional analysis of the effects Ric-8A has on G-protein alpha subunit guanine nucleotide-binding activities. The use of Ric-8A to prepare GTPgammaS-Galpha and nucleotide-free Galpha (in complex with Ric-8A) is described.
Department of Pharmacology, University of Texas Southwestern Medical Center, Dallas 75390, USA.
Gregory G
GG
Tall
Alfred G
AG
Gilman
D000818:Animals; D002845:Chromatography / Q000379:methods; D020558:GTP Phosphohydrolases / Q000378:metabolism; D044385:GTP-Binding Protein alpha Subunits / Q000378:metabolism*; D020662:Guanine Nucleotide Exchange Factors / Q000235:genetics / Q000302:isolation & purification* / Q000378:metabolism*; D016244:Guanosine 5'-O-(3-Thiotriphosphate) / Q000302:isolation & purification / Q000378:metabolism; D006160:Guanosine Triphosphate / Q000378:metabolism; D009687:Nuclear Proteins / Q000235:genetics / Q000302:isolation & purification* / Q000378:metabolism*; D051381:Rats; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D015398:Signal Transduction / Q000502:physiology
D044385:GTP-Binding Protein alpha Subunits; D020662:Guanine Nucleotide Exchange Factors; D009687:Nuclear Proteins; D011993:Recombinant Fusion Proteins; C485885:Ric8a protein, rat; D016244:Guanosine 5'-O-(3-Thiotriphosphate); D006160:Guanosine Triphosphate; D020558:GTP Phosphohydrolases