title: Mechanisms and interaction of vinblastine and reduced glutathione transport in membrane vesicles by the rabbit multidrug resistance protein Mrp2 expressed in insect cells.
abstract: The present study examined how the multidrug resistance protein (MRP) 2, which is an ATP-dependent anionic conjugate transporter, also mediates transport of the chemotherapeutic cationic drug vinblastine (VBL). We show that ATP-dependent [(3)H]VBL (0.2 microM) uptake into membrane vesicles from Sf9 cells infected with a baculovirus encoding rabbit Mrp2 (Sf9-Mrp2) was similar to vesicles from mock-infected Sf9 cells (Sf9-mock) but could be stimulated by reduced glutathione (GSH) with a half-maximum stimulation of 1.9 +/- 0.1 mM. At 5 mM GSH, initial ATP-dependent [(3)H]VBL uptake rates were saturable with an apparent K(m) of 1.5 +/- 0.3 microM. The inhibitory effect of VBL on Mrp2-mediated ATP-dependent transport of the anionic conjugate [(3)H]leukotriene C(4) was potentiated by increasing GSH concentrations. Membrane vesicles from Sf9-Mrp2 cells exhibited a approximately 7-fold increase in initial GSH uptake rates compared with membrane vesicles from Sf9-mock cells. Uptake of [(3)H]GSH was osmotically sensitive, independent of ATP, and was trans-inhibited by GSH. The anionic conjugates estradiol-17beta-D-glucuronide and leukotriene C(4) cis-inhibited [(3)H]GSH uptake but only in the presence of ATP. Whereas ATP-dependent [(3)H]VBL uptake was stimulated by GSH, VBL did not affect [(3)H]GSH uptake. Our results show that GSH is required for Mrp2-mediated ATP-dependent VBL transport and that Mrp2 transports GSH independent of VBL.
authors.affiliation: Department of Pharmacology and Toxicology, University of Nijmegen, The Netherlands.
authors.forename: R A
authors.initials: RA
authors.lastname: Van Aubel
authors.forename: J B
authors.initials: JB
authors.lastname: Koenderink
authors.forename: J G
authors.initials: JG
authors.lastname: Peters
authors.forename: C H
authors.initials: CH
authors.lastname: Van Os
authors.forename: F G
authors.initials: FG
authors.lastname: Russel
mesh_terms: D000818:Animals; D000972:Antineoplastic Agents, Phytogenic / Q000378:metabolism*; D001692:Biological Transport; D002460:Cell Line; D005978:Glutathione / Q000378:metabolism*; D007313:Insecta; D017997:Leukotriene C4 / Q000493:pharmacokinetics; D024101:Mitochondrial Proteins; D011817:Rabbits; D012269:Ribosomal Proteins / Q000235:genetics / Q000378:metabolism*; D029701:Saccharomyces cerevisiae Proteins; D014162:Transfection; D014316:Tritium; D014747:Vinblastine / Q000378:metabolism*
chemical_list: D000972:Antineoplastic Agents, Phytogenic; C093947:MRP2 protein, S cerevisiae; D024101:Mitochondrial Proteins; D012269:Ribosomal Proteins; D029701:Saccharomyces cerevisiae Proteins; D014316:Tritium; D017997:Leukotriene C4; D014747:Vinblastine; D005978:Glutathione