[Isolation of GTP-binding Gs-protein liberated from human erythrocyte membranes under the action of fluoride ions].
Human erythrocyte membranes were incubated in the presence of sodium fluoride. After centrifugation at 30,000 g for 30 min the supernatant was able to stimulate the catalytic subunit of adenylate cyclase. The stimulatory factor was purified from the supernatant of fluoride-treated membranes by three subsequent chromatographic steps including DEAE-Sephacel ion-exchange chromatography in the absence of detergent, gel-filtration on Ultrogel AcA 44 in the presence of 1% sodium cholate and phenyl-Sepharose CL/4B hydrophobic chromatography. The final preparation showed approximately 120-fold purification in stimulatory activity over the initial extract and contained two polypeptides (Mr 42 kDa and 36 kDa). The stimulator activity of the preparation was inhibited by 60% by beta gamma-subunits of the GTP-binding protein of bovine brain membranes, G0. The data obtained suggest that the regulatory GTP-binding stimulatory protein of adenylate cyclase, GS, dissociates from human erythrocyte membranes as a result of fluoride-ion treatment.
Skurat
A V
AV
Iurkova
M S
MS
Kots
A Ia
AIa
Bulargina
T V
TV
D000262:Adenylyl Cyclases / Q000378:metabolism; D000818:Animals; D001921:Brain / Q000378:metabolism; D002417:Cattle; D002853:Chromatography, Liquid; D004591:Electrophoresis, Polyacrylamide Gel; D004789:Enzyme Activation; D004910:Erythrocyte Membrane / Q000187:drug effects / Q000378:metabolism*; D005459:Fluorides / Q000494:pharmacology*; D019204:GTP-Binding Proteins / Q000302:isolation & purification* / Q000378:metabolism; D006801:Humans; D007454:Iodides / Q000494:pharmacology*
D007454:Iodides; D019204:GTP-Binding Proteins; D000262:Adenylyl Cyclases; D005459:Fluorides
Vydelenie reguliatornogo GTP-sviayvaiushchego Gs-belka, osvobozhdaiushchegosia iz membran éritrotsitov cheloveka pod deĭstviem ftorid-iona.