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  A simple liposomal system to reconstitute and assay highly efficient Na+/D-glucose cotransport from kidney brush-border membranes

Ducis, I., & Koepsell, H. (1983). A simple liposomal system to reconstitute and assay highly efficient Na+/D-glucose cotransport from kidney brush-border membranes. Biochimica et Biophysica Acta-Biomembranes, 730(1), 119-129. doi:10.1016/0005-2736(83)90324-3.

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 Creators:
Ducis, Ilze1, Author           
Koepsell, Hermann1, Author           
Affiliations:
1Department of Physiology, Max Planck Institute of Biophysics, Max Planck Society, ou_2068297              

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Free keywords: Na+; Glucose transport; Cotransport; Liposome; Membrane reconstitution; Lipid-protein interaction; (Pig kidney)
 Abstract: A simple procedure to reconstitute highly efficient Na+/D-glucose cotransport from solubilized brush-border membranes of proximal kidney tubules is described. Reconstitution of transport activity was possible with various phospholipid and cholesterol combinations; the presence, however, of cholesterol and at least one phospholipid was essential. When liposomes were synthesized from only one phospholipid and cholesterol, the highest uptake rats were observed with phosphatidylserine; phosphatidylcholine was less effective and phosphatidylethanolamine showed insignificant uptake of D-glucose in the presence of Na+. The rate at which an inward-directed Na+ gradient dissipated across the liposomal membranes was reduced if the cholesterol concentration of liposomes was increased. In the optimized system, proteoliposomes were formed from cholesterol and phosphatidylserine by a heat-sonication-freeze-thaw procedure. A Na+-gradient persisted for hours across these proteoliposomal membranes and a Na+/D-glucose cotransport with the following characteristics could be demonstrated: (1) dependency on the Na+ gradient; (2) a transient (3) rheogenicity; (4) stereospecificity; and (5) high-affinity phlorizin inhibition. Since the Na+-gradient-stimulated D-glucose uptake is linear for minutes, the initial uptake rates can be measured and the Na+/D-glucose cotransport activity of different protein fractions can be compared.

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Language(s): eng - English
 Dates: 1982-10-182003-01-301983-04-21
 Publication Status: Issued
 Pages: 11
 Publishing info: -
 Table of Contents: -
 Rev. Type: Peer
 Identifiers: DOI: 10.1016/0005-2736(83)90324-3
PMID: 6681984
 Degree: -

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Title: Biochimica et Biophysica Acta-Biomembranes
Source Genre: Journal
 Creator(s):
Affiliations:
Publ. Info: Amsterdam : Elsevier
Pages: - Volume / Issue: 730 (1) Sequence Number: - Start / End Page: 119 - 129 Identifier: ISSN: 0005-2736
CoNE: https://pure.mpg.de/cone/journals/resource/954926938702