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

C. Özvegy-Laczka, Thomas Litman, G. Szakács, Zoltán Nagy, Susan Bates, A. Váradi, B. Sarkadi

Research output: Contribution to journalArticle

245 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 - 2001

Fingerprint

Insects
Adenosine Triphosphatases
Sf9 Cells
Rhodamine 123
Mitoxantrone
Proteins
Ethylmaleimide
Daunorubicin
ATP-Binding Cassette Transporters
Vanadates
Prazosin
Multiple Drug Resistance
Cell membranes
Verapamil
Drug Resistance
Doxorubicin
Cyclosporine
Neoplasms
Cell Membrane
Pharmacology

Keywords

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

ASJC Scopus subject areas

  • Biochemistry
  • Biophysics
  • Molecular Biology

Cite this

Functional characterization of the human multidrug transporter, ABCG2, expressed in insect cells. / Özvegy-Laczka, C.; Litman, Thomas; Szakács, G.; Nagy, Zoltán; Bates, Susan; Váradi, A.; Sarkadi, B.

In: Biochemical and Biophysical Research Communications, Vol. 285, No. 1, 2001, p. 111-117.

Research output: Contribution to journalArticle

@article{b72055097f1b4b7bb5174533a772a122,
title = "Functional characterization of the human multidrug transporter, ABCG2, expressed in insect cells",
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.",
keywords = "ABC half-transporter, ABCG2, BCRP, Drug-stimulated ATPase activity, Multidrug resistance, MXR, Sf9 cells",
author = "C. {\"O}zvegy-Laczka and Thomas Litman and G. Szak{\'a}cs and Zolt{\'a}n Nagy and Susan Bates and A. V{\'a}radi and B. Sarkadi",
year = "2001",
doi = "10.1006/bbrc.2001.5130",
language = "English",
volume = "285",
pages = "111--117",
journal = "Biochemical and Biophysical Research Communications",
issn = "0006-291X",
publisher = "Academic Press Inc.",
number = "1",

}

TY - JOUR

T1 - Functional characterization of the human multidrug transporter, ABCG2, expressed in insect cells

AU - Özvegy-Laczka, C.

AU - Litman, Thomas

AU - Szakács, G.

AU - Nagy, Zoltán

AU - Bates, Susan

AU - Váradi, A.

AU - Sarkadi, B.

PY - 2001

Y1 - 2001

N2 - 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.

AB - 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.

KW - ABC half-transporter

KW - ABCG2

KW - BCRP

KW - Drug-stimulated ATPase activity

KW - Multidrug resistance

KW - MXR

KW - Sf9 cells

UR - http://www.scopus.com/inward/record.url?scp=0034789387&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0034789387&partnerID=8YFLogxK

U2 - 10.1006/bbrc.2001.5130

DO - 10.1006/bbrc.2001.5130

M3 - Article

C2 - 11437380

AN - SCOPUS:0034789387

VL - 285

SP - 111

EP - 117

JO - Biochemical and Biophysical Research Communications

JF - Biochemical and Biophysical Research Communications

SN - 0006-291X

IS - 1

ER -