Redox modulation of the response of NADH oxidase activity of rat liver plasma membranes to cyclic AMP plus ATP.

NADH oxidase activity of rat liver plasma membranes was inhibited by low concentrations (1-100 nM) of ATP. The inhibition was amplified by addition of nanomolar concentrations (0.1-10) of cyclic AMP. The inhibition was complex and related to a marked increase in the Km for NADH at high NADH concentrations together with a concomitant decrease in the Vmax. In the absence of added or residual ATP, cyclic AMP was without effect. The response of cyclic AMP + ATP was inhibited by low concentrations of the selective inhibitor of cyclic AMP-dependent protein kinase, H-89 but not by staurosporin. The Vmax but not the Km was modified by treating the plasma membranes with a mild oxidizing agent, N-chlorosuccinamide, or with the reducing agent, dithiothreitol. In the presence of dithiothreitol, the Vmax was reduced by cyclic AMP + ATP. In contrast, in the presence of N-chlorosuccinamide, the Vmax was increased by cyclic AMP + ATP relative to cyclic AMP + ATP alone. Thus, the effect of cyclic AMP + ATP on the Vmax could be either an increase or a decrease depending on whether the membranes were oxidized or reduced. The results demonstrate regulation of NADH oxidase activity of rat liver plasma membranes through cyclic AMP-mediated phosphorylation by membrane-located protein kinase activities where the final response is dependent on the oxidation-reduction status of the plasma membranes.