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Biochemistry, Biophysics, and Structural Biology

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2009

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Ligand-Dependent Effects On The Conformational Equilibrium Of The Na+,K+-Atpase As Monitored By Voltage Clamp Fluorometry, Robert Dempski, Stefan Geys, Ernst Bamberg Jun 2009

Ligand-Dependent Effects On The Conformational Equilibrium Of The Na+,K+-Atpase As Monitored By Voltage Clamp Fluorometry, Robert Dempski, Stefan Geys, Ernst Bamberg

Robert E. Dempski

Voltage clamp fluorometry was used to monitor conformational changes associated with electrogenic partial reactions of the Na(+),K(+)-ATPase after changes in the concentration of internal sodium (Na(+)(i)) or external potassium (K(+)(o)). To probe the effects of the Na(+)(i) concentration on the Na(+) branch of the Na(+),K(+)-ATPase, oocytes were depleted of Na(+)(i) and then loaded with external sodium (Na(+)(o)) using the amiloride-sensitive epithelial sodium channel. The K(+) branch of the Na(+),K(+)-ATPase was studied by exposing the oocytes to different K(+)(o) concentrations in the presence and absence of Na(+)(o) to obtain additional information on the apparent affinity for K(+)(o). Our results demonstrate that …