Multidrug resistance protein mrp2 mediates ATP-dependent transport of classic renal organic anion p-aminohippurate.
p-Aminohippurate (PAH) is widely used as a model substrate to characterize organic anion transport in kidney proximal tubules. The carrier responsible for uptake of PAH across the basolateral membrane has been cloned and well characterized, whereas transporters mediating PAH excretion across the brush-border (apical) membrane are yet unknown. In this study we investigated whether PAH is a substrate for the apical multidrug resistance protein 2 (Mrp2). Overexpression of recombinant rabbit Mrp2 in Sf9 cells significantly increased ATP-dependent [(14)C]PAH uptake into isolated membrane vesicles compared with endogenous ATP-dependent uptake. The Michaelis-Menten constant and maximal velocity for Mrp2-mediated ATP-dependent [(14)C]PAH transport were 1.9 +/- 0.8 mM and 187 +/- 29 pmol. mg(-1). min(-1), respectively. On the basis of the inhibitory profile, the endogenous ATP-dependent PAH transporter does not appear to be an ortholog of Mrp2. Together, our results show that Mrp2 is a low-affinity ATP-dependent PAH transporter, indicating that Mrp2 might contribute to urinary PAH excretion.
Van Aubel
R A
RA
Department of Pharmacology and Toxicology, Faculty of Medical Sciences, University Medical Centre Nijmegen, 6500 HB Nijmegen, The Netherlands.
Peters
J G
JG
Masereeuw
R
R
Van Os
C H
CH
Russel
F G
FG
D018435:ATP Binding Cassette Transporter, Subfamily B / Q000502:physiology*; D000255:Adenosine Triphosphate / Q000502:physiology*; D000818:Animals; D000838:Anions / Q000378:metabolism; D001692:Biological Transport; D002352:Carrier Proteins / Q000378:metabolism; D002460:Cell Line; D007668:Kidney / Q000378:metabolism*; D007700:Kinetics; D026901:Membrane Transport Proteins; D000089762:Multidrug Resistance-Associated Protein 2; D027425:ATP-Binding Cassette, Sub-Family C Proteins; D011817:Rabbits; D011994:Recombinant Proteins / Q000378:metabolism; D018411:Spodoptera / Q000166:cytology; D013997:Time Factors; D010130:p-Aminohippuric Acid / Q000378:metabolism*
D018435:ATP Binding Cassette Transporter, Subfamily B; D000255:Adenosine Triphosphate; D000838:Anions; D002352:Carrier Proteins; D026901:Membrane Transport Proteins; D000089762:Multidrug Resistance-Associated Protein 2; D027425:ATP-Binding Cassette, Sub-Family C Proteins; D011994:Recombinant Proteins; D010130:p-Aminohippuric Acid; C421109:multidrug resistance-associated protein 1