Access channel model for the voltage dependence of the forward-running Na+/K+ pump.

The voltage dependence of steady state current produced by the forward mode of operation of the endogenous electrogenic Na+/K+ pump in Na(+)-loaded Xenopus oocytes has been examined using a two-microelectrode voltage clamp technique. Four experimental cases (in a total of 18 different experimental conditions) were explored: variation of external Na+ (Nao) at saturating (10 mM) external K+ (Ko), and activation of pump current by various Ko at 0, 15, and 120 mM Nao (tetramethylammonium replacement). Ionic current through K+ channels was blocked by Ba2+ (5 mM) and tetraethylammonium (20 mM), thereby allowing pump-mediated current to be measured by addition or removal of external K+. Control measurements and corrections were made for pump current run-down and holding current drift. Additional controls were done to estimate the magnitude of the inwardly directed pump-mediated current that was present in K(+)-free solution and the residual K(+)-channel current. A pseudo two-state access channel model is described in the Appendix in which only the pseudo first-order rate coefficients for binding of external Na+ and K+ are assumed to be voltage dependent and all transitions between states in the Na+/K+ pump cycle are assumed to be voltage independent. Any three-state or higher order model with only two oppositely directed voltage-dependent rate coefficients can be reduced to an equivalent pseudo two-state model. The steady state current-voltage (I-V) equations derived from the model for each case were simultaneously fit to the I-V data for all four experimental cases and yielded least-squares estimates of the model parameters. The apparent fractional depth of the external access channel for Na+ is 0.486 +/- 0.010; for K+ it is 0.256 +/- 0.009. The Hill coefficient for Na+ is 2.18 +/- 0.06, and the Hill coefficient for K+ (which is dependent on Nao) ranges from 0.581 +/- 0.019 to 1.35 +/- 0.034 for 0 and 120 mM Nao, respectively. The model provides a reasonable fit to the data and supports the hypothesis that under conditions of saturating internal Na+, the principal voltage dependence of the Na+/K+ pump cycle is a consequence of the existence of an external high-field access channel in the pump molecule through which Na+ and K+ ions must pass in order to reach their binding sites.