Rapid purification and characterization of F1-ATPase of Vibrio parahaemolyticus.
The F1 portion of H(+)-translocating ATPase as purified from membrane vesicles of Vibrio parahaemolyticus by a rapid procedure. The whole purification process (from culture of cells to purification of the enzyme) could be completed in 1 day. The F1-ATPase consists of five subunits (alpha, beta, gamma, delta and epsilon) like F1 of Escherichia coli and other microorganisms. The F1-ATPase of V. parahaemolyticus showed some interesting properties. Its activity was greatly stimulated by high concentrations (about 0.5 M) of SO4(2-), SO3(2-) and CH3COO-, their effects decreasing in this order. Among the anions tested, Cl- and NO3- were ineffective, or rather inhibitory, and cations had no significant effects. Ethanol (or methanol) stimulated the activity 2- to 3-fold. The activity was inhibited by 4-acetamido-4'-isothiocyanostilbene 2,2'-disulfonate (SITS) (an anion exchanger inhibitor), tetrachlorosalicylanilide (TCS) (an H+ conductor), azide and N-ethylmaleimide. Zinc inhibited the activity only slightly, although it strongly inhibited the ATPase activity in membrane vesicles.
Sakai
Y
Y
Department of Microbiology, Faculty of Pharmaceutical Sciences, Okayama, Japan.
Kanazawa
H
H
Tsuda
M
M
Tsuchiya
T
T
D012856:4-Acetamido-4'-isothiocyanatostilbene-2,2'-disulfonic Acid / Q000494:pharmacology; D000838:Anions; D001386:Azides / Q000494:pharmacology; D002413:Cations, Divalent; D002462:Cell Membrane / Q000201:enzymology; D002845:Chromatography; D004591:Electrophoresis, Polyacrylamide Gel; D004789:Enzyme Activation / Q000187:drug effects; D004926:Escherichia coli / Q000201:enzymology; D005033:Ethylmaleimide / Q000494:pharmacology; D008274:Magnesium / Q000494:pharmacology; D006180:Proton-Translocating ATPases / Q000037:antagonists & inhibitors / Q000302:isolation & purification* / Q000378:metabolism; D012458:Salicylanilides / Q000494:pharmacology; D014736:Vibrio parahaemolyticus / Q000201:enzymology*
D000838:Anions; D001386:Azides; D002413:Cations, Divalent; D012458:Salicylanilides; D012856:4-Acetamido-4'-isothiocyanatostilbene-2,2'-disulfonic Acid; D006180:Proton-Translocating ATPases; C003390:3,3',4',5-tetrachlorosalicylanilide; D008274:Magnesium; D005033:Ethylmaleimide