The specificity of omeprazole as an (H+ + K+)-ATPase inhibitor depends upon the means of its activation.

Omeprazole (OME) is a novel acid secretion inhibitor, believed to act directly on the gastric proton pump, the (H+ + K+)-ATPase. Inhibition of ATPase activity is associated with an incorporation of [14C]OME into gastric vesicles containing the (H+ + K+)-ATPase, and both processes are greatly enhanced if the OME is exposed to acidic pH. This, and other evidence, suggests that the acidic environment of the (H+ + K+)-ATPase generates from OME a reactive intermediate which covalently inhibits the pump. We have compared the means by which the OME was acid-activated with the specificity of inhibition (amount of incorporation of omeprazole required to produce 100% inhibition of K+-stimulated ATPase activity). The stoichiometry of incorporation has been related to the number of detectable catalytic phosphorylation sites in each preparation (an index of the number of functional pumps). In lyophilised gastric vesicles, where the membrane barriers separating the cytoplasmic and luminal faces of the enzyme are substantially destroyed, incubation with OME at pH 6.1 produced a progressive inhibition and incorporation over 120 min. Complete inhibition of K+-ATPase required 13 +/- 3 (SEM; N = 4) moles of OME incorporation per phosphorylation site. In intact gastric vesicles, under conditions shown independently to result in proton pumping and the acidification of the vesicle interior (150 mM KCl, 9 microM valinomycin, 2 mM Mg-ATP pH 7.0), inhibition and incorporation occurred more rapidly (15 min). Complete inhibition of K+-ATPase required only 1.8 +/- 0.15 (SEM; N = 3) moles of OME per phosphorylation site.(ABSTRACT TRUNCATED AT 250 WORDS)