TY - JOUR T1 - Adenosine Triphosphate-Dependent Transport of Anionic Conjugates by the Rabbit Multidrug Resistance-Associated Protein Mrp2 Expressed in Insect Cells JF - Molecular Pharmacology JO - Mol Pharmacol SP - 1062 LP - 1067 VL - 53 IS - 6 AU - Rémon A. M. H. van Aubel AU - Marcel A. van Kuijck AU - Jan B. Koenderink AU - Peter M. T. Deen AU - Carel H. van Os AU - Frans G. M. Russel Y1 - 1998/06/01 UR - http://molpharm.aspetjournals.org/content/53/6/1062.abstract N2 - The multidrug resistance-associated protein Mrp2 is expressed in liver, kidney, and small intestine and mediates ATP-dependent transport of conjugated organic anions across the apical membrane of epithelial cells. We recently cloned a rabbit cDNA encoding a protein that on basis of highest amino acid homology and tissue distribution was considered to be the rabbit homolog of rat Mrp2. To investigate whether rabbit Mrp2 mediates ATP-dependent transport similar to rat Mrp2, we expressed rabbit Mrp2 in Spodoptera frugiperda (Sf9) cells using recombinant baculovirus. Mrp2 was expressed as an underglycosylated protein in Sf9 cells and to a higher level compared with rabbit liver and renal proximal tubules. Both 17β-estradiol-17-β-d-glucuronide ([3H]E217βG, 50 nm) and [3H]leukotriene C4 (3 nm) were taken up by Sf9-Mrp2 membrane vesicles in an ATP-dependent fashion. Uptake of [3H]E217βG was dependent on the osmolarity of the medium and saturable for ATP (Km = 623 μm). Leukotriene C4, MK571, phenolphthalein glucuronide, and fluorescein-methotrexate were good inhibitors of [3H]E217βG transport. The inhibitory potency of cyclosporin A and methotrexate was moderate, whereas fluorescein, α-naphthyl-β-d-glucuronide, andp-nitrophenyl-β-d-glucuronide did not inhibit transport. In conclusion, we show direct ATP-dependent transport by recombinant rabbit Mrp2 and provide new data on Mrp2 inhibitor specificity. The American Society for Pharmacology and Experimental Therapeutics ER -