Na+,K(+)-ATPase pump currents activated by an ATP concentration jump. Comparison of studies with purified membrane fragments and giant excised patches.

Abstract

The lipid bilayer and the giant patch technique were used to study the Na+,K(+)-ATPase. In excised patches from ventricular myocytes, Na+,K(+)-pump currents show a saturable ATP dependence with a Km of approximately 150 microM at 24 degrees C. Partial reactions in the transport cycle were investigated by generating ATP concentration jumps through photolytic release of ATP from caged ATP at pH 7.4 and 6.3. Transient outward currents were obtained at pH 6.3 with a fast rising phase followed by a slow decay to a stationary current. Experiments with purified pig kidney Na+,K(+)-ATPase attached to a planar lipid bilayer resulted in similar pump current signals and the same outcome. It was concluded that the fast rate constant of approximately 200 s-1 at 24 degrees C reflects a step rate limiting the electrogenic Na+ release.