Rapid kinetic analyses of the Na+/K+-ATPase distinguish among different criteria for conformational change

Pratap, Promod R.; Robinson, Joseph D.

doi:10.1016/0005-2736(93)90075-b

Cited by 40 publications

(47 citation statements)

References 31 publications

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“…This dye has a very fast response to the changes in the membrane potential and has been used to study the presteady state kinetics of the Na ϩ -K ϩ -ATPase (35)(36)(37)(38)(39). The purified Na ϩ -NQR was reconstituted into proteoliposomes, following the protocol described before (14).…”

Section: Methodsmentioning

confidence: 99%

The Kinetic Reaction Mechanism of the Vibrio cholerae Sodium-dependent NADH Dehydrogenase

Tuz

Mezic

et al. 2015

Journal of Biological Chemistry

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Section: Methodsmentioning

confidence: 99%

The Kinetic Reaction Mechanism of the Vibrio cholerae Sodium-dependent NADH Dehydrogenase

Tuz

Mezic

et al. 2015

Journal of Biological Chemistry

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“…The ®rst approach made use of direct electric measurements, either by electrophysiological techniques applied to native cells (Rakowski et al, 1990, Gadsby, Rakowski & De Weer, 1993Rakowski, 1993;Hilgemann, 1994;Holmgren et al, 2000) or by use of capacitive coupling of open membrane fragments puri®ed from kidney tissue, which were adsorbed tightly to planar lipid bilayers (Fendler et al, 1985;Borlinghaus, Apell & LaÈ uger, 1987;Wuddel & Apell, 1995;Sokolov et al, 1998;Pintschovius, Fendler & Bamberg, 1999;Domaszewicz & Apell, 1999). As an alternative method an indirect approach was introduced, which utilizes electrochromic styryl dyes that allow the detection of charge movements in the Na,K-ATPase (Klodos & Forbush, III, 1988;BuÈ hler et al, 1991;StuÈ rmer et al, 199l;Pratap & Robinson, 1993;Heyse et al, 1994;Fedosova, Cornelius & Klodos, 1995;Klodos, Fedosova & Cornelius, 1997;Clarke et al, 1998;Schneeberger & Apell, 1999). In the case of the CaATPase of the sarcoplasmatic reticulum (SR) the situation is di erent: direct electric measurements are not possible since the membrane of the SR is extremely leaky for monovalent cations, a property that short-circuits both sides of the membrane.…”

Section: Introductionmentioning

confidence: 99%

Detection of Charge Movements in Ion Pumps by a Family of Styryl Dyes

Pedersen

Roudná

Beutner

et al. 2002

Journal of Membrane Biology

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Abstract. A family of¯uorescent styryl dyes was synthesized to apply them as probes that monitor the ion-translocating activity of the Na,K-ATPase and the SR Ca-ATPase, similar to the widely used dye RH421. All dyes had the same chromophore but they di ered in the length of the spacer between chromophore and polar head, an isothiocyanate group, and in the lengths of the two identical acyl chains, which form the tail of the dye molecules. A number of substrate-dependent partial reactions of both P-type ATPases a ected the¯uorescence intensity, and the magnitude of the¯uorescence changes was used to characterize the usefulness of the dyes for further application. The experimental results indicate that electrochromy is the major mechanism of these dyes. While in the case of the Na,K-ATPase a single dye, 5QITC, showed larger¯uorescence changes than all others, in the case of the SR Ca-ATPase all dyes tested were almost equal in their¯uorescence responses. This prominent di erence is interpreted as a hint that the position of the ion binding sites in both ion pumps may di er signi®cantly despite their otherwise closely related structural features. Quench experiments with spin-labeled lipids in various positions of their fatty acids were used to gain information on the depth of the chromophore of the di erent dyes within the membrane dielectric, however, the spatial resolution was so poor that only qualitative information on the position of the chromophore in the lipid phase could be obtained.

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“…Thiol-reactive derivatives of fluorescein, rhodamine, and pyrene are commercially available reagents used to investigate the structurefunction relationships of proteins like the Na ϩ -K ϩ ATPase (16,17) and F-actin (18). Of these probes, pyrene derivatives are the longest lived with lifetimes near 100 ns.…”

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confidence: 99%