title: Human organic anion transporter OAT1 is not responsible for glutathione transport but mediates transport of glutamate derivatives.
abstract: Due to their clearance function, the kidneys are exposed to high concentrations of oxidants and potentially toxic substances. To maintain cellular integrity, renal cells have to be protected by sufficient concentrations of the antioxidant glutathione (GSH). We tested whether GSH or its precursors are taken up by human organic anion transporters 1 (OAT1) and 3 (OAT3) stably expressed in HEK293 cells. GSH did not inhibit uptake of p-aminohippurate (PAH) or of estrone sulfate (ES) in OAT3-transfected HEK293 cells. In OAT1-transfected cells, GSH reduced the uptake of PAH marginally. Among the GSH constituent amino acids, glutamate, cysteine, and glycine, only glutamate inhibited OAT1, but labeled glutamate was not taken up by a probenecid-inhibitable transport system. Thus OAT1 binds glutamate but is unable to translocate it. The GSH precursor dipeptide, cysteinyl glycine (cysgly), and the glutamate derivative N-acetyl glutamate (NAG), inhibited uptake of PAH when present in the medium and trans-stimulated uptake of PAH from the intracellular side, indicating that they are hitherto unrecognized transported substrates of OAT1. N-acetyl aspartate weakly interacted with OAT1, but aspartate did not. NAG inhibited also OAT3, albeit with much lower affinity compared with OAT1, and glutamate did not interact with OAT3 at all. Taken together, human OAT3 and OAT1 cannot be involved in renal GSH extraction from the blood. However, OAT1 could support intracellular GSH synthesis by taking up cysteinyl glycine.
authors.affiliation: Department of Systemic Physiology and Pathophysiology, University Medical Center Göttingen, Göttingen, Germany.
authors.forename: Yohannes
authors.initials: Y
authors.lastname: Hagos
authors.forename: Gerhard
authors.initials: G
authors.lastname: Burckhardt
authors.forename: Birgitta C
authors.initials: BC
authors.lastname: Burckhardt
mesh_terms: D000618:Aminohippuric Acids / Q000378:metabolism; D001224:Aspartic Acid / Q000031:analogs & derivatives / Q000378:metabolism; D001692:Biological Transport; D003545:Cysteine / Q000378:metabolism; D004151:Dipeptides / Q000378:metabolism; D004970:Estrone / Q000031:analogs & derivatives / Q000378:metabolism; D005971:Glutamates / Q000378:metabolism*; D005978:Glutathione / Q000378:metabolism*; D005998:Glycine / Q000378:metabolism; D057809:HEK293 Cells; D006801:Humans; D007668:Kidney / Q000187:drug effects / Q000378:metabolism; D027362:Organic Anion Transport Protein 1 / Q000235:genetics / Q000378:metabolism*; D029362:Organic Anion Transporters, Sodium-Independent / Q000235:genetics / Q000378:metabolism; D011339:Probenecid / Q000494:pharmacology; D014162:Transfection; D014528:Uricosuric Agents / Q000494:pharmacology
chemical_list: D000618:Aminohippuric Acids; D004151:Dipeptides; D005971:Glutamates; D027362:Organic Anion Transport Protein 1; D029362:Organic Anion Transporters, Sodium-Independent; D014528:Uricosuric Agents; C114338:organic anion transport protein 3; D004970:Estrone; D001224:Aspartic Acid; C028505:cysteinylglycine; D005978:Glutathione; D003545:Cysteine; C016195:N-acetylglutamic acid; D011339:Probenecid; C017296:estrone sulfate; D005998:Glycine