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Drug-resistant tumor cells actively extrude a variety of chemotherapeutic agents by the action of the multi-drug resistance (MDR1) gene product, the plasma membrane P-glycoprotein. In this report we show that the expression of the human MDR1 gene in cultured Sf9 insect cells via a baculovirus vector generates a high activity vanadate-sensitive membrane ATPase. This ATPase is markedly stimulated by drugs known to interact with the P-glycoprotein, such as vinblastine and verapamil, and the ability of the various drugs to stimulate the ATPase corresponds to their previously observed affinity for this transporter. The drug-stimulated ATPase is not present in uninfected or mock-infected Sf9 cells, and its appearance correlates with the appearance of the MDR1 gene product detected with a monoclonal anti-MDR protein antibody and by labeling with 8-azido-ATP. The drug-induced ATPase requires magnesium ions, does not utilize ADP or AMP as substrates, exhibits a half-maximal activation at about 0.5 mM MgATP, and its maximal activity (about 3-5 mumol/mg MDR protein/min) approaches that of the well characterized ion transport ATPases. These results provide the first direct demonstration of a high capacity drug-stimulated ATPase activity of the human multidrug resistance protein and offer a new and simple assay for the investigation of functional interactions of various drugs with this clinically important enzyme.
B Sarkadi, and
E M Price, and
R C Boucher, and
U A Germann, and
G A Scarborough
November 2006,
FEBS letters,
B Sarkadi, and
E M Price, and
R C Boucher, and
U A Germann, and
G A Scarborough
January 1996,
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B Sarkadi, and
E M Price, and
R C Boucher, and
U A Germann, and
G A Scarborough
November 1995,
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B Sarkadi, and
E M Price, and
R C Boucher, and
U A Germann, and
G A Scarborough
January 1998,
Methods in enzymology,
B Sarkadi, and
E M Price, and
R C Boucher, and
U A Germann, and
G A Scarborough
September 1997,
The Journal of biological chemistry,
B Sarkadi, and
E M Price, and
R C Boucher, and
U A Germann, and
G A Scarborough
January 1998,
Sheng wu hua xue yu sheng wu wu li xue bao Acta biochimica et biophysica Sinica,
B Sarkadi, and
E M Price, and
R C Boucher, and
U A Germann, and
G A Scarborough
February 1995,
Journal of bioenergetics and biomembranes,
B Sarkadi, and
E M Price, and
R C Boucher, and
U A Germann, and
G A Scarborough
November 1999,
Biochimica et biophysica acta,
B Sarkadi, and
E M Price, and
R C Boucher, and
U A Germann, and
G A Scarborough
December 1993,
Shi yan sheng wu xue bao,
B Sarkadi, and
E M Price, and
R C Boucher, and
U A Germann, and
G A Scarborough
July 1995,
Blood,
