Combined localization and real-time functional studies using a GFP-tagged ABCG2 multidrug transporter

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16 Citations (Scopus)

Abstract

ABCG2 is a half-transporter which causes multidrug resistance when overexpressed in tumor cells. Availability of combined localization and functional assays would greatly improve cell biology and drug modulation studies for this transporter. Here we demonstrate that an N-terminally GFP-tagged version of the protein (GFP-G2) can be used to directly monitor ABCG2 expression, dimerization, localization and function in living cells. GFP-G2 is fully functional when tested for drug-stimulated ATPase activity, vesicular transport assay, subcellular localization or cell surface epitope conformational changes. By measuring both GFP and Hoechst 33342 dye fluorescence in HEK-293 cells, we provide evidence that a real-time transport assay can be reliably applied to identify ABCG2 substrates, transport modulators, as well as to monitor the cellular functions of this multidrug transporter protein. This approach also avoids the need of cloning, drug selection or other further separation or characterization of the transgene-expressing cells.

Original languageEnglish
Pages (from-to)667-673
Number of pages7
JournalBiochemical and biophysical research communications
Volume367
Issue number3
DOIs
Publication statusPublished - Mar 14 2008

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Keywords

  • ABCG2
  • Dye transport
  • Functional assay
  • Green fluorescent protein
  • Hoechst 33342
  • Multidrug resistance

ASJC Scopus subject areas

  • Biophysics
  • Biochemistry
  • Molecular Biology
  • Cell Biology

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