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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 A; Ismair, Manfred G; Kubitz, Ralf; Schmitt, Marcus; Häussinger, Dieter; Stieger, Bruno; Hagenbuch, Bruno; Meier, Peter J; Beuers, Ulrich; Paumgartner, Gustav (2000). Stable expression and functional characterization of a Na+-taurocholate cotransporting green fluorescent protein in human hepatoblastoma HepG2 cells. Cytotechnology, 34(1/2):1-9.

Abstract

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

Abstract

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

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Item Type:Journal Article, refereed, further contribution
Communities & Collections:National licences > 142-005
Dewey Decimal Classification:Unspecified
Language:English
Date:1 January 2000
Deposited On:26 Sep 2018 12:47
Last Modified:24 Nov 2018 03:08
Publisher:Springer
ISSN:0920-9069
OA Status:Green
Free access at:Publisher DOI. An embargo period may apply.
Publisher DOI:https://doi.org/10.1023/a:1008152729133
Related URLs:https://www.swissbib.ch/Search/Results?lookfor=nationallicencespringer101023A1008152729133 (Library Catalogue)

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