Reactivity of gastric (H+ + K+)-ATPase to N-ethoxycarbonyl-2-ethoxy-1,2-dihydroquinoline.

The K+-dependent ATPase and p-nitrophenyl phosphatase activity of, and formation of phosphoenzyme by, hog parietal cell membranes were inhibited in a time- and concentration-dependent manner by the carboxyl-activating reagent, N-ethoxycarbonyl-2-ethoxy-1,2-dihydroquinoline (EEDQ). The kinetics of inactivation was pseudo first order and was similar to the EEDQ-catalyzed incorporation of [14C]glycine ethyl ester. The most likely mechanism is the EEDQ-dependent formation of inter- and intramolecular amide bonds. Cross-linking between the subunits of the ATPase occurs with EEDQ treatment. The presence of K+ on the luminal face of the enzyme is able to prevent EEDQ inhibition of K+ ATPase activity (but not intermolecular cross-linking), whereas ATP enhanced the rate of inactivation. EEDQ reaction with the enzyme therefore allows investigation of K+- and ATP-dependent states of the enzyme.