Fast transient currents in Na,K-ATPase induced by ATP concentration jumps from the P3-[1-(3',5'-Dimethoxyphenyl)-2-Phenyl-2-Oxo]ethyl ester of ATP
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Electrogenic ion transport by Na,K-ATPase was investigated by analysis of transient currents in a model system of protein-containing membrane fragments adsorbed to planar lipid bilayers. Sodium transport was triggered by ATP concentration jumps in which ATP was released from an inactive precursor by an intense near-UV light flash. The method has been used previously with the P3-1-(2-nitrophenyl)ethyl ester of ATP (NPE-caged ATP), from which the relatively slow rate of ATP release limits analysis of processes in the pump mechanism controlled by rate constants greater than 100 s1 at physiological pH. Here Na,K-ATPase was reinvestigated using the P3-[1-(3,5-dimethoxyphenyl)-2-phenyl-2-oxo]ethyl ester of ATP (DMB-caged ATP), which has an ATP release rate of >105 s1. Under otherwise identical conditions, photorelease of ATP from DMB-caged ATP showed faster kinetics of the transient current compared to that from NPE-caged ATP. With DMB-caged ATP, transient currents had rate profiles that were relatively insensitive to pH and the concentration of caged compound. Rate constants of ATP binding and of the E1 to E2 conformational change were compatible with earlier studies. Rate constants of enzyme phosphorylation and ADP-dependent dephosphorylation were 600 s1 and 1.5 × 106 M1 s1, respectively, at pH 7.2 and 22°C.
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SOKOLOV, Valerij S., Hans-Jürgen APELL, John E. T. CORRIE, David R. TRENTHAM, 1998. Fast transient currents in Na,K-ATPase induced by ATP concentration jumps from the P3-[1-(3',5'-Dimethoxyphenyl)-2-Phenyl-2-Oxo]ethyl ester of ATP. In: Biophysical Journal. 1998, 74(5), pp. 2285-2298. ISSN 0006-3495. Available under: doi: 10.1016/S0006-3495(98)77938-XBibTex
@article{Sokolov1998trans-8419, year={1998}, doi={10.1016/S0006-3495(98)77938-X}, title={Fast transient currents in Na,K-ATPase induced by ATP concentration jumps from the P3-[1-(3',5'-Dimethoxyphenyl)-2-Phenyl-2-Oxo]ethyl ester of ATP}, number={5}, volume={74}, issn={0006-3495}, journal={Biophysical Journal}, pages={2285--2298}, author={Sokolov, Valerij S. and Apell, Hans-Jürgen and Corrie, John E. T. and Trentham, David R.} }
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