Heterologous expression and functional characterization of a mouse renal organic anion transporter in mammalian cells.

Article Details

Citation

Kuze K, Graves P, Leahy A, Wilson P, Stuhlmann H, You G

Heterologous expression and functional characterization of a mouse renal organic anion transporter in mammalian cells.

J Biol Chem. 1999 Jan 15;274(3):1519-24.

PubMed ID
9880528 [ View in PubMed
]
Abstract

Organic anion transporters play an essential role in eliminating a wide range of organic anions including endogenous compounds, xenobiotics, and their metabolites from kidney, thereby preventing their potentially toxic effects within the body. The goal of this study was to extend our previous study on the functional characterization and post-translational modification of a mouse kidney organic anion transporter (mOAT), in a mammalian cell system, COS-7 cells. The transporter-mediated p-aminohippurate (PAH) uptake was saturable, probenecid-sensitive, and inhibited by a wide range of organic anions including vitamins, anti-hypertensive drugs, anti-tumor drugs, and anti-inflammatory drugs. Tunicamycin, an inhibitor of asparagine-linked glycosylation, significantly inhibited the transport activity. Immunofluorescence provided evidence that most of the protein remained in the intracellular compartment in tunicamycin-treated cells. Diethyl pyrocarbonate (DEPC), a histidine residue-specific reagent, completely blocked PAH transport. The inhibitory effect by DEPC was significantly protected (90%) by pretreating the cells with excess unlabeled PAH, suggesting that the histidine residues may be close to the PAH binding sites. Finally, in situ mRNA localization was studied in postnatal mouse kidney. The expression was observed in proximal tubules throughout development. We conclude that COS-7 cells may be useful in pharmacological and molecular biological studies of this carrier. The carbohydrate moieties are necessary for the proper trafficking of mOAT to the plasma membrane, and histidine residues appear to be important for the transport function.

DrugBank Data that Cites this Article

Drug Transporters
DrugTransporterKindOrganismPharmacological ActionActions
Aminohippuric acidSolute carrier family 22 member 6ProteinHumans
Unknown
Substrate
Inhibitor
Details
CaptoprilSolute carrier family 22 member 6ProteinHumans
Unknown
Substrate
Inhibitor
Details
CarprofenSolute carrier family 22 member 6ProteinHumans
Unknown
Inhibitor
Details
DiclofenacSolute carrier family 22 member 6ProteinHumans
Unknown
Inhibitor
Details
EnalaprilSolute carrier family 22 member 6ProteinHumans
Unknown
Inhibitor
Details
Folic acidSolute carrier family 22 member 6ProteinHumans
Unknown
Inhibitor
Details
IndomethacinSolute carrier family 22 member 6ProteinHumans
Unknown
Substrate
Inhibitor
Details
MethotrexateSolute carrier family 22 member 6ProteinHumans
Unknown
Substrate
Inhibitor
Details
ProbenecidSolute carrier family 22 member 6ProteinHumans
Unknown
Substrate
Inhibitor
Details
RiboflavinSolute carrier family 22 member 6ProteinHumans
Unknown
Inhibitor
Details
SulindacSolute carrier family 22 member 6ProteinHumans
Unknown
Inhibitor
Details