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Preparation of sodium potassium atpase ec 3.6.1.3 containing liposomes with predictable transport properties by a procedure relating the sodium potassium transport capacity to the atpase activity


, : Preparation of sodium potassium atpase ec 3.6.1.3 containing liposomes with predictable transport properties by a procedure relating the sodium potassium transport capacity to the atpase activity. Journal of Biochemical and Biophysical Methods 5(6): 299-306

A microprocedure for the preparation of Na,K-ATPase-containing liposomes with a minimal starting material (200 .mu.g) of purified Na,K-ATPase is presented. Phosphatidylcholine is added gradually to cholate-solubilized Na,K-ATPase of various concentrations and the lipid-induced decrease in enzyme activity is monitored. After removal of the detergent by dialysis, the transport parameters of the resulting Na,K-ATPase-liposomes are established by a microassay. By relating the transport properties to the Na,K-ATPase activity present before dialysis, a procedure is developed which allows to prepare standardized Na,K-ATPase-liposomes with predictable transport properties.


Accession: 006173879

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Related references

Anner, B.M.; Moosmayer, M., 1982: Preparation of Na,K-ATPase-containing liposomes with predictable transport properties by a procedure relating the Na,K-transport capacity to the ATPase activity. A microprocedure for the preparation of Na,K-ATPase-containing liposomes with a minimal starting material (200 microgram) of purified Na,K-ATPase is presented. Phosphatidylcholine is added gradually to cholate-solubilized Na,K-ATPase of various co...

Jorgensen P.L.; Anner B.M., 1979: Purification and characterization of sodium ion plus potassium ion stimulated atpase part 8 altered sodium ion to potassium ion transport ratio in vesicles reconstituted with purified sodium ion plus potassium ion stimulated atpase that has been selectively modified with trypsin in presence of sodium chloride. Using the cholate dialysis technique for reconstitution of (Na+ + K+)-ATPase into phospholipid vesicles, the transport properties of purified (Na+ + K+)-ATPase from the [pig] outer renal medulla were examined after selective modification of its pr...

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Berberian G.; Beauge L., 1992: Phosphatase activity and potassium transport in liposomes with sodium potassium atpase incorporated. We have used liposomes with incorporated pig kidney Na+,K+-ATPase to study vanadate sensitive K+-K+ exchange and net K+ uptake under conditions of acetyl- and p-nitrophenyl phosphatase activities. The experiments were performed at 20.degree. C. Cy...

Hokin L.E.; Dixon J.F., 1979: Parameters of reconstituted sodium and potassium transport in liposomes in which purified sodium potassium atpase is incorporated by freeze thaw sonication. Skou, J C And J G Norby (Ed ) Na,k-Atpase: Structure And Kinetics; 2nd International Conference on Properties And Functions Of Na,k-Atpase, Sonderborg, Denmark, Sept 17-21, 1978 Xii+549p Academic Press, Inc : New York, N Y , Usa Illus P47-68

Anner, B.M., 1981: A transport model for the sodium potassium atpase ec 3.6.1.3 in liposomes including the sodium potassium channel function. (Na+-K+)ATPase liposomes of various degrees of reconstitution are formed by varying the amount of phosphatidylcholine added to the soluble [rabbit kidney] (Na+K+)ATPase before vesicles are formed by cholate removal. In the presence of ATP the reco...

Anner, B.M.; Ting-Beall, H.P.; Robertson, J.D., 1984: Characterization of sodium potassium atpase liposomes 2. effect of alpha subunit digestion on intra membrane particle formation and sodium potassium transport. The effect of the protein structure of [rabbit kidney] (Na+ + K+)-ATPase on its incorporation into liposome membranes was investigated. The catalytic .alpha.-subunit of (Na+ + K+)-ATPase was split into low-MW fragments by trypsin treatment and the...

Hara Y.; Yamada J.; Nakao M., 1986: Proton transport catalyzed by the sodium pump ouabain sensitive atpase activity and the phosphorylation of sodium potassium atpase in the absence of sodium ions. The possibility that H+ might substitute for Na+ at Na+ sites of Na+,K+-ATPase was studied. Na+,K+-ATPase purified from pig kidney showed ouabain-sensitive K+-dependent ATPase activity in the absence of Na+ at acid pH (H+,K+-ATPase). The specific...

Anonymous, 1973: Purification and some properties of sodium and potassium transport atpase ec 3613 part 3 comparison of lipid and protein components of sodium and potassium atpase preparations at various purification steps from pig brain

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