Influence of the salts of choline on the rate limiting step of active transport of sodium ions by Na,K-ATPase

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406 First publ. in: Biologicheskie Membrany 21 (2004), pp. 406-414

Konstanzer Online-Publikations-System (KOPS) URL: http://www.ub.uni-konstanz.de/kops/volltexte/2007/4162/

URN: http://nbn-resolving.de/urn:nbn:de:bsz:352-opus-41628

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Influence of the Choline Salts of on the Rate Limiting Step of Active Transport of Sodium Ions by Na+, K+-ATPase

A. G. Ayuyan

¹^,

V. S. Sokolov

¹^,

H.-J. Apel1

²

IFrumkin Institute of Electrochemistry, Russian Academy of Sciences, 31 Leninski Prospect, Moscow, 117071 , Russia;

2Department ofBiology, University of Konstanz, Fach M635, 78457 Konstanz, Germany

The influence of temperature and concentrated salts of choline with different anions on the kinetics of

electrogenic Na+ transport by the Na+, K+-ATPase through membrane fragments adsorbed on BLMs was

studied. The kinetic parameters of the transport were determined from the short circuit current caused by

fast release of ATP from Caged-ATP due to UV light flash. The temperature dependency of the time con-

stant

't2

of the exponential decay of short circuit current allowed the determination of its apparent activa-

BHOJIOrHQECKHE MEMBPAHbI

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tion energy (according to equation 1/'t2 = A . exp(-Ea/Rn) of the rate-limiting step of transport, that is a conformational transition of Na+, K+-ATPase. Iodide or bromide ions decreased the activation energy, E

_{a ,}

and also the pre-exponential factor, A, of the temperature dependency. The effect of A was dominant, lead- ing to decreased rate constants. The different effects of these salts on two sides of the protein were ob- served. It was shown that the salts were considerably more effective when they were present on both (ex- tracellular and cytoplasmic) sides of protein compared to the case when they were only at cytoplasmic side. It means that these salts effectively influence the functioning of the pump at the extracellular side only. The effect of these salts can be explained by its influence as chaotropic ions on the energy of the protein surface contacting with water. Side sensitiveness can be due to different changes of contact areas of water and protein on both sides of the membrane due to conformational transition, that decreases the cytoplasmic surface and increases the extracellular one. The effect of choline chloride was quite different than that of iodide or bromide. It did not change the activation energy and was more effective at cytoplasmic side of the membrane. This can be explained by asymmetric screening of negatively charged membrane frag- ment with Na+, K+-ATPase that leads to rise of transmembrane potential influencing transport of sodium ions by protein.

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