Differential regulation of voltage-gated [K.sup.+] channels by oxidized and reduced pyridine nucleotide coenzymes

The activity of the voltage-sensitive [K.sup.+] (Kv) channels varies as a function of the intracellular redox state and metabolism, and several Kv channels act as oxygen sensors. However, the mechanisms underlying the metabolic and redox regulation of these channels remain unclear. In this study we investigated the regulation of Kv channels by pyridine nucleotides. Heterologous expression of Kv[alpha] 1.5 in COS-7 cells led to the appearance of noninactivating currents. Inclusion of 0.1-1 mM NA[D.sup.+] or 0.03-0.5 mM NAD[P.sup.+] in the internal solution of the patch pipette did not affect Kv currents. However, 0.5 and 1 mM NA[D.sup.+] and 0.1 and 0.5 mM NAD[P.sup.+] prevented inactivation of Kv currents in cells transfected with Kv[alpaha] 1.5 and Kv[beta] 1.3 and shifted the voltage dependence of activation to depolarized potentials. The Kv[beta]-dependent inactivation of Kv[alpha] currents was also decreased by internal pipette perfusion of the cell with 1 mM NA[D.sup.+]. The Kv[alpha]1.5-Kv[beta]1.3 currents were unaffected by the internal application of 0.1 mM NADPH or 0.1 or 1 mM NADH. Excised inside-out patches from cells expressing Kv[alpha]1.5-Kv[beta] 1.3 showed transient single-channel activity. The mean open time and the open probability of these currents were increased by the inclusion of 1 mM NA[D.sup.+] in the perfusate. These results suggest that NAD[(P).sup.+] prevents Kv[beta]-mediated inactivation of Kv currents and provide a novel mechanism by which pyridine nucleotides could regulate specific [K.sup.+] currents as a function of the cellular redox state INAD(P)H-to-NAD[(P).sup.+] ratio]. Shaker potassium ion channels: Kv[beta] subunits: patch clamp: aldo-keto reductase: COS-7 cells