Functional characterization of the human multidrug transporter, ABCG2, expressed in insect cells

Csilla Özvegy, Thomas Litman, Gergely Szakács, Zoltán Nagy, Susan Bates, András Váradi, Balázs Sarkadi

Research output: Contribution to journalArticle

246 Citations (Scopus)

Abstract

ABCG2 (also called MXR (3), BCRP (4), or ABCP (5)) is a recently-identified ABC half-transporter, which causes multidrug resistance in cancer. Here we report that the expression of the ABCG2 protein in Sf9 insect cells resulted in a high-capacity, vanadate-sensitive ATPase activity in isolated membrane preparations. ABCG2 was expressed underglycosylated, and its ATPase activity was stimulated by daunorubicin, doxorubicin, mitoxantrone, prazosin and rhodamine 123, compounds known to be transported by this protein. ABCG2-ATPase was inhibited by low concentrations of Na-orthovanadate, N-ethylmaleimide and cyclosporin A. Verapamil had no effect, while Fumitremorgin C, reversing ABCG2-dependent cancer drug resistance, strongly inhibited this ATPase activity. The functional expression of ABCG2 in this heterologous system indicates that no additional partner protein is required for the activity of this multidrug transporter, probably working as a homodimer. We suggest that the Sf9 cell membrane ATPase system is an efficient tool for examining the interactions of ABCG2 with pharmacological agents.

Original languageEnglish
Pages (from-to)111-117
Number of pages7
JournalBiochemical and Biophysical Research Communications
Volume285
Issue number1
DOIs
Publication statusPublished - Jan 1 2001

Keywords

  • ABC half-transporter
  • ABCG2
  • BCRP
  • Drug-stimulated ATPase activity
  • MXR
  • Multidrug resistance
  • Sf9 cells

ASJC Scopus subject areas

  • Biophysics
  • Biochemistry
  • Molecular Biology
  • Cell Biology

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