Simulation of the kinetics of adenosine 5′-triphosphate hydrolysis catalyzed by the Cu2+ ion: The role of conformation and the catalytic effect of the OH− ion

Abstract

The hydrolysis kinetics of the dimeric complex (CuATP2− · OH2)2 {D} up to ≈40% ATP conversion at 25°C, pH 5.7–7.8, and [Cu · ATP]0 = (2.07 ± 0.03) × 10−3 mol/l is analyzed by numerical simulation. CuADP− + Pi (Pi is an inorganic phosphate) form from DOH−, and the latter forms rapidly from D. The abstraction of H+ from the coordinated H2O molecule is an irreversible reaction involving an OH− ion from the medium. The maximum possible DOH− concentration at a given pH is reached at the initial stage of hydrolysis (0.3–6.0 min after the initiation of hydrolysis). CuADP− + Pi form from D via two consecutive irreversible steps. The ADP buildup rate in the process is determined by the reversible conformational transformation of DOH− resulting in a pentacovalent intermediate (IntK). OH− ions from the medium are involved both in IntK formation and in the reverse reaction and are a hydrolysis inhibitor. AMP forms from the intermediate IntK3, which forms reversibly from DOH−, OH− ions from the medium being involved in the forward and reverse reactions. This is followed by irreversible (AMPH)− formation involving H3O+ ions from the medium. The rate and equilibrium constants are determined for the formation and decomposition of hydrolysis intermediates. The concentrations of the intermediates are plotted versus time for various pH values. The structures of the intermediates are suggested. The causes of a peak appearing in the initial ADP formation rate versus pH curve are analyzed.