Separation and characterization of Na+,K+ -ATPase containing vesicles

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Separation_and_Characterization.pdf
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1994
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Ivashchuk-Kienbaum, Yu. A.
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Biochimica et Biophysica Acta / Biomembranes. 1994, 1196(1), pp. 29-37. ISSN 0005-2736. Available under: doi: 10.1016/0005-2736(94)90291-7
Zusammenfassung

Na/K-ATPase was reconstituted in vesicles prepared by a dialysis method. Ion-exchange chromatography was used to obtain well characterized fractions from the inhomogeneous vesicle preparation. Lipid and protein content wasdetermined by optical methods during the elution process. It was possible to separate fractions with distinct enzymatic and transport activities. A protocol was set up, which allowed to calculate the average number of 5-IAF labeled ion pumps per vesicle in the different fractiuns. The dependence of the number of protein molecules per vesicle was studied as function of the initial protein concentration added to the lipid solution before dialysis. The transport activity disappears completely at very low protein concentrations (3.3 mg protein per mg lipid). This observation is in favor of the proposal discllssed in the literature, that the hcterodimer (alpha-beta)2 is the transport-active form of the Na,K -ATPase. The presented method can be applied to all reconstituted vesicle preparations in which the proteins can be labeled quantitatively with a fluorescence dye.

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Fachgebiet (DDC)
570 Biowissenschaften, Biologie
Schlagwörter
ATPase, Na/K-, Proteoliposomes, Ion-exchange chromatography, Fluorescence labeling, Electrogenic ion transport
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ISO 690IVASHCHUK-KIENBAUM, Yu. A., Hans-Jürgen APELL, 1994. Separation and characterization of Na+,K+ -ATPase containing vesicles. In: Biochimica et Biophysica Acta / Biomembranes. 1994, 1196(1), pp. 29-37. ISSN 0005-2736. Available under: doi: 10.1016/0005-2736(94)90291-7
BibTex
@article{IvashchukKienbaum1994Separ-8771,
  year={1994},
  doi={10.1016/0005-2736(94)90291-7},
  title={Separation and characterization of Na+,K+ -ATPase containing vesicles},
  number={1},
  volume={1196},
  issn={0005-2736},
  journal={Biochimica et Biophysica Acta / Biomembranes},
  pages={29--37},
  author={Ivashchuk-Kienbaum, Yu. A. and Apell, Hans-Jürgen}
}
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