Catalysis of oxygen-18 exchange between inorganic phosphate and water by the gastric H,K-ATPase.
The gastric H,K-ATPase is shown to catalyze 18O exchange between Pi and HOH. Mg2+ is the only ion required for the reaction. K+ increases the rate of isotope exchange, which is directly proportional to specific ATPase activity. Ouabain, which potently inhibits the Na,K-ATPase, has no effect on the exchange reaction. Conversely, omeprazole, which is specific for the H,K-ATPase, completely inhibits 18O exchange. Vanadate inhibition of exchange can be explained by competitive binding with Pi. The rate of 18O exchange is faster than the hydrolytic rate and about equal to the dephosphorylation rate. Thus, the ionic requirements for exchange, inhibition of exchange, and the rate of exchange are all compatible with catalysis occurring via the same phosphoenzyme intermediate formed during hydrolysis of ATP. The distribution of 18O-labeled Pi species formed with time indicates that Pi loss is only about twice as fast as covalent bond formation. This kinetic pattern is unaffected by K+, temperature, or the specific activity of the enzyme preparation. Invariance of the kinetic pattern could mean isotope exchange is always catalyzed by the same form of the enzyme, and K+ and higher temperature accelerate the reaction by increasing the relative amount of the active conformer. Independence of the kinetic pattern from specific activity implies that the catalytic mechanism of active enzyme molecules is unaffected by inactive proteins in gastric microsomal membranes.
D000251:Adenosine Triphosphatases / Q000037:antagonists & inhibitors / Q000378:metabolism*; D000818:Animals; D001562:Benzimidazoles / Q000494:pharmacology; D005753:Gastric Mucosa / Q000201:enzymology*; D017506:H(+)-K(+)-Exchanging ATPase; D066298:In Vitro Techniques; D007477:Ions; D007700:Kinetics; D009853:Omeprazole; D010103:Oxygen Isotopes; D010710:Phosphates; D013552:Swine; D014867:Water
D001562:Benzimidazoles; D007477:Ions; D010103:Oxygen Isotopes; D010710:Phosphates; D014867:Water; D000251:Adenosine Triphosphatases; D017506:H(+)-K(+)-Exchanging ATPase; D009853:Omeprazole