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

Biochemical and Biophysical Research Communications
Tamás I OrbánL Homolya

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.

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Citations

Nov 6, 2012·The Journal of Biological Chemistry·Tufan GökirmakAmro Hamdoun
Jan 13, 2009·Advanced Drug Delivery Reviews·Csilla HegedusBalázs Sarkadi
Mar 28, 2012·Biochemical and Biophysical Research Communications·Csilla HegedüsBalázs Sarkadi
Aug 10, 2010·Biochimica Et Biophysica Acta·László HomolyaBalázs Sarkadi
Jun 22, 2012·Molecular Membrane Biology·Christian RoosFrank Bernhard
Jul 17, 2014·Drug Discovery Today. Technologies·Elena L RudashevskayaPeter Chiba
May 27, 2016·PloS One·Zoltán Hegyi, László Homolya
Sep 23, 2010·Drug Metabolism and Disposition : the Biological Fate of Chemicals·Yuki IkebuchiHiroshi Suzuki
Jan 14, 2020·Xenobiotica; the Fate of Foreign Compounds in Biological Systems·Maryam Yousaf, Moazzam Ali
Feb 27, 2021·Frontiers in Cell and Developmental Biology·Zsuzsa Bartos, László Homolya
Apr 4, 2021·International Journal of Molecular Sciences·László Homolya

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