RAPID PURIFICATION AND CHARACTERIZATION OF F1-ATPASE OF VIBRIO-PARAHAEMOLYTICUS

The F1 portion of H+-translocating ATPase was 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 (α, β, γ, δ and ε{lunate}) 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 SO2-4, SO2-3 and CH3COO-, their effects decreasing in this order. Among the anions tested, Cl- and NO-3 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. © 1990.