Expression of the human multidrug resistance cDNA in insect cells generates a high activity drug-stimulated membrane ATPase.

These results provide the first direct demonstration of a high capacity drug-stimulated ATPase activity of the human multidrug resistance proteinand offer a new and simple assay for the investigationof functional interactions of various drugs with thicslinically important enzyme

B Sarkadi; E.M. Price; R.C. Boucher; U.A. Germann; G.A. Scarborough

2021

Scholarcy highlights

  • Drug-resistant tumorcells actively extrudea variety coded by the MDRl gene, and the expression of the MDRl of chemotherapeutic agents by the actionof the multi- cDNA in drug-sensitive cells has been demonstrated to confer drug resistance gene product, the plasma a multidrug resistance phenotype
  • The drug-stimtransporters and with mammalian proteins such as the cystic fibrosis transmembrane conductance regulator.The MDR protein consists of 1280amino acids and has been proposed to contain 12 membrane-spanning hydrophobic domains, two nucleotide-binding domains, and specific N-linked glycosylation sites. Certain aspects of this proposed transmembrane topography and a tisulated ATPase is not present in uninfected or mock- sue-specific, variable glycosylation which probably does not infected Sf9 cells, and its appearance correlates with affect the multidrug transporter function have been confirmed the appearance of the MDRl gene product detected by biochemical approaches
  • 3-5 pmol/mg MDR protein/min) approachesthat 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 proteinand offer a new and simple assay for the investigationof functional interactions of various drugs with thicslinically important enzyme
  • The protein binds ATP, andmutagenesis of the proposed nucleotide-binding domains has been shown to result in a failure of the expressed protein to confer drug resistance. All these data collectively suggest that the MDR gene product is a membrane ATPase which functions as an active drug transporter
  • Inthis report we demonstrate that the expression of the MDRl gene in Sf9 insect cells resultsin the appearance of a drug-stimulated ATPase activity with a high specific activity approaching that of the ion-transporting ATPases
  • Thedatainthis paperdemonstrate the presence of a heretofore undetected large capacity, drug-stimulated, vanadate-sensitive membrane ATPase activity directly related to the presence of the human MDRl gene product

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