Cloning of the pig renal organic anion transporter 1 (pOAT1).
A pig kidney cDNA library was screened for the porcine ortholog of the multispecific organic anion transporter 1 (pOAT1). Several positive clones were isolated resulting in two alternatively spliced cDNA clones of pOAT1 (pOAT1 and pOAT1A). pOAT1-cDNAs consist of 2126 or 1895 base pairs (EMBL Acc. No. AJ308234 and AJ308235) encoding 547 or 533 amino acid residue proteins with 89, 87, 83 and 81% homology to the human, rabbit, rat, and mouse OAT1, respectively. Heterologous expression of pOAT1 in Xenopus laevis oocytes revealed an apparent K(m) for [3H]PAH of 3.75 +/- 1.6 microM. [3H]PAH uptake mediated by pOAT1 was abolished by 0.5 mM glutarate or 1 mM probenecid. Functional characterization of pOAT1A did not show any affinity for [3H]PAH. In summary, we cloned two alternative splice variants of the pig ortholog of organic anion transporter 1. One splice form (pOAT1) showed typical functional characteristics of organic anion transporter 1, whereas the second form appears not to transport PAH.
Hagos
Yohannes
Y
Zentrum für Physiologie und Pathophysiologie, Abt Vegetative Physiologie, Universität Göttingen, Humboldtallee 23, 37073 Göttingen, Germany. hagos@veg-physiol.med.uni-goettingen.de
Bahn
Andrew
A
Asif
Abdul R
AR
Krick
Wolfgang
W
Sendler
Mark
M
Burckhardt
Gerhard
G
D017398:Alternative Splicing; D000595:Amino Acid Sequence; D000818:Animals; D000838:Anions / Q000378:metabolism; D002352:Carrier Proteins / Q000235:genetics / Q000378:metabolism; D003001:Cloning, Molecular; D017124:Conserved Sequence / Q000235:genetics; D018076:DNA, Complementary / Q000737:chemistry / Q000235:genetics; D015723:Gene Library; D007668:Kidney / Q000378:metabolism*; D007700:Kinetics; D008969:Molecular Sequence Data; D009865:Oocytes / Q000502:physiology; D027362:Organic Anion Transport Protein 1 / Q000235:genetics*; D020033:Protein Isoforms / Q000235:genetics / Q000378:metabolism; D017422:Sequence Analysis, DNA; D017386:Sequence Homology, Amino Acid; D013552:Swine / Q000235:genetics*; D014982:Xenopus laevis; D010130:p-Aminohippuric Acid / Q000493:pharmacokinetics
D000838:Anions; D002352:Carrier Proteins; D018076:DNA, Complementary; D027362:Organic Anion Transport Protein 1; D020033:Protein Isoforms; D010130:p-Aminohippuric Acid