Identification and functional assessment of the novel murine organic anion transporter Oat5 (Slc22a19) expressed in kidney.
An uncharacterized murine cDNA clone was identified and, through sequence, phylogenetic, and functional analysis, determined to encode the newest member of the organic anion transporter family, organic anion transporter 5 (Oat5; Slc22a19). The Oat5 cDNA clone contained an insert 1,964 bp in length with a predicted open reading frame (from bp 84 to bp 1,739) coding for a peptide 551 amino acids long. Slc22a19 was localized to mouse chromosome 19 near the genes encoding Oat1 (Slc22a6) and Oat3 (Slc22a8). Northern blot analysis revealed Oat5 is highly expressed in the kidney of adult mice and rats. No sexual dimorphism in renal or hepatic expression of Oat5 was observed. Unlike Oat1-3, Oat5 expression was not detected in the choroid plexus of either mice or rats. Murine Oat5-expressing Xenopus laevis oocytes supported increased accumulation of the mycotoxin ochratoxin A, compared with water-injected control oocytes. This uptake was significantly inhibited by probenecid and the organic anions 2,4-dichlorophenoxyacetic acid, salicylate, and estrone sulfate but not by para-aminohippurate or urate. Transport of ochratoxin A by murine Oat5 was saturable, with an estimated K(m) of 2.0 +/- 0.45 microM. Oat5-mediated transport was neither cis-inhibited nor trans-stimulated by the dicarboxylate glutarate. Uptake was also completely unaffected by short-circuiting of the membrane potential. Thus the motive forces behind Oat5 function, which provide insight into its membrane localization, need to be further resolved. These data demonstrate for the first time that this newly identified gene encodes a protein that functions as an organic anion transporter.
Youngblood
Geri L
GL
Department of Pharmaceutical Sciences, Medical University of South Carolina, Charleston, SC 29425, USA.
Sweet
Douglas H
DH
D015084:2,4-Dichlorophenoxyacetic Acid / Q000494:pharmacology; D000595:Amino Acid Sequence; D000818:Animals; D004305:Dose-Response Relationship, Drug; D004563:Electrochemistry; D004970:Estrone / Q000031:analogs & derivatives* / Q000494:pharmacology; D005260:Female; D005977:Glutarates / Q000494:pharmacology; D007668:Kidney / Q000378:metabolism*; D051379:Mice; D008969:Molecular Sequence Data; D009183:Mycotoxins / Q000493:pharmacokinetics; D009793:Ochratoxins / Q000493:pharmacokinetics; D009865:Oocytes; D027361:Organic Anion Transporters / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism / Q000502:physiology*; D011188:Potassium / Q000008:administration & dosage; D012333:RNA, Messenger / Q000378:metabolism; D051381:Rats; D012459:Salicylates / Q000494:pharmacology; D014018:Tissue Distribution; D014982:Xenopus laevis
D005977:Glutarates; D009183:Mycotoxins; D009793:Ochratoxins; D027361:Organic Anion Transporters; D012333:RNA, Messenger; D012459:Salicylates; C488753:Slc22a19 protein, mouse; C025589:ochratoxin A; D015084:2,4-Dichlorophenoxyacetic Acid; D004970:Estrone; C017296:estrone sulfate; D011188:Potassium