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Stable expression and functional characterization of a Na+-taurocholate cotransporting green fluorescent protein in human hepatoblastoma HepG2 cells

Kullak-Ublick, Gerd ; Ismair, Manfred ; Kubitz, Ralf ; Schmitt, Marcus ; Häussinger, Dieter ; Stieger, Bruno ; Hagenbuch, Bruno ; Meier, Peter ; Beuers, Ulrich ; Paumgartner, Gustav

In: Cytotechnology, 2000, vol. 34, no. 1-2, p. 1-9

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    Titel
    • Stable expression and functional characterization of a Na+-taurocholate cotransporting green fluorescent protein in human hepatoblastoma HepG2 cells
    Autor
    • Kullak-Ublick, Gerd. Divisions of Clinical Pharmacology/Toxicology, Department of Medicine, University Hospital, CH-8091, Zürich, Switzerland
    • Ismair, Manfred. Divisions of Clinical Pharmacology/Toxicology, Department of Medicine, University Hospital, CH-8091, Zürich, Switzerland
    • Kubitz, Ralf. Department of Medicine, University of Düsseldorf, D-40225, Düsseldorf, Germany
    • Schmitt, Marcus. Department of Medicine, University of Düsseldorf, D-40225, Düsseldorf, Germany
    • Häussinger, Dieter. Department of Medicine, University of Düsseldorf, D-40225, Düsseldorf, Germany
    • Stieger, Bruno. Divisions of Clinical Pharmacology/Toxicology, Department of Medicine, University Hospital, CH-8091, Zürich, Switzerland
    • Hagenbuch, Bruno. Divisions of Clinical Pharmacology/Toxicology, Department of Medicine, University Hospital, CH-8091, Zürich, Switzerland
    • Meier, Peter. Divisions of Clinical Pharmacology/Toxicology, Department of Medicine, University Hospital, CH-8091, Zürich, Switzerland
    • Beuers, Ulrich. Department of Medicine II, Klinikum Grosshadern, University of Munich, D-81377, Munich, Germany
    • Paumgartner, Gustav. Department of Medicine II, Klinikum Grosshadern, University of Munich, D-81377, Munich, Germany
    Dokumententyp
    • Postprint
    Sprache
    • Englisch
    Veröffentlicht in
    • Cytotechnology, 2000, vol. 34, no. 1-2, p. 1-9. Kluwer Academic Publishers
    Andere elektronische Ausgabe
    OAI-PMH-ID
    • oai:doc.rero.ch:311324
    Summary
    Sodium-dependent uptake of bile acids from blood is aliver-specific function which is mediated by theNa+-taurocholate cotransporting polypeptide(Ntcp). We report the stable expression of aNa+-taurocholate cotransporting green fluorescentfusion protein in the human hepatoblastoma cell lineHepG2, normally lacking Ntcp expression. Ntcp-EGFPassociated green fluorescence colocalized with Ntcpimmunofluorescence in the plasma membrane. Intransfected HepG2 cells, the fusion protein mediatedthe sodium-dependent uptake of the bile acidtaurocholate (Km: 24.6 μmol/l) and of the anionicsteroids estrone-3-sulfate and dehydroepiandrosteronesulfate. We conclude that the Ntcp-EGFP fusion proteinfollows the sorting route of Ntcp, is functionallyidentical to Ntcp and could be used to monitor proteintrafficking in living HepG2 cells