Effect of chaotropic anions on the sodium transport by the Na,K-ATPase.
The effect of choline iodide, bromide and chloride on the kinetics of the electrogenic sodium transport by the Na,K-ATPase was investigated in a model system of ATPase-containing membrane fragments adsorbed on the lipid bilayer membrane. The kinetic parameters of Na(+) transport were determined from short circuit currents after fast release of ATP from its caged precursor. The falling phase of the current transients could be fitted by a single exponential with the time constant, tau (2). Its temperature dependence allowed an estimation of the activation energy of the rate-limiting reaction step, the conformation transition E(1)/E(2). Choline iodide and bromide caused a decrease of the activation energy as well as the overall rate of the process expressed as the pre-exponential factor A of the Arrhenius equation. If choline iodide or bromide were present on the cytoplasmic and extracellular sides of the protein, the temperature dependent changes were more pronounced than when present on the cytoplasmic side only. These results can be explained by an effect of the anions on water structure on the extracellular surface of the protein, where a deep access channel connects the ion-binding sites with the solution. Chloride ions also caused a deceleration of the electrogenic transport, however, in contrast to iodide or bromide, they did not affect the activation energy, and were more effective when added on the cytoplasmic side. This effect can be explained by asymmetric screening of the negative surface charges which leads to a transmembrane electric potential that modifies the ion transfer.
Ayuyan
Artem G
AG
A.N. Frumkin Institute of Physical Chemistry and Electrochemistry, Russian Academy of Sciences, 31 Leninski Prospect, 119071 Moscow, Russia.
Sokolov
Valerij S
VS
Lenz
Alexander A
AA
Apell
Hans-Jürgen
HJ
D000255:Adenosine Triphosphate / Q000378:metabolism; D000818:Animals; D000838:Anions / Q000378:metabolism*; D001665:Binding Sites; D001965:Bromides / Q000378:metabolism; D002712:Chlorides / Q000378:metabolism; D002794:Choline / Q000378:metabolism; D006868:Hydrolysis; D007454:Iodides / Q000378:metabolism; D017136:Ion Transport; D007700:Kinetics; D008051:Lipid Bilayers / Q000378:metabolism; D011485:Protein Binding; D011487:Protein Conformation; D011817:Rabbits; D012964:Sodium / Q000378:metabolism*; D000254:Sodium-Potassium-Exchanging ATPase / Q000378:metabolism*; D012996:Solutions; D055672:Static Electricity; D013696:Temperature; D014867:Water / Q000737:chemistry
D000838:Anions; D001965:Bromides; D002712:Chlorides; D007454:Iodides; D008051:Lipid Bilayers; D012996:Solutions; D014867:Water; D000255:Adenosine Triphosphate; D012964:Sodium; D000254:Sodium-Potassium-Exchanging ATPase; D002794:Choline