Molecular cloning and functional characterization of the bovine (Bos taurus) organic anion transporting polypeptide Oatp1a2 (Slco1a2).
We describe the cloning, functional characterization and tissue localization of a novel membrane transporter of the OATP/Oatp-gene family obtained from liver and kidney of cattle (Bos taurus). The carrier protein exhibits highest sequence identity to the human OATP1A2 (previously called OATP-A) and is, therefore, named bovine Oatp1a2. Bovine Oatp1a2 received the gene symbol Slco1a2 that is identical to the SLC classification of human OATP1A2 (SLCO1A2, previously called SLC21A3) and is likely an orthologue of the human gene. Two different full-length bOatp1a2 cDNAs of 2316-bp and 3504-bp were obtained and encoded for a 666 amino acid membrane protein, which contains twelve putative transmembrane spanning domains. Bovine Oatp1a2 expression was detected in liver, kidney, brain and adrenal gland. Uptake studies in cRNA-injected oocytes demonstrated that bOatp1a2 transports estrone-3-sulfate and taurocholate, with K(m) values of 9.6 microM and 51 microM, respectively, and estradiol-17beta-glucuronide. However, the structurally-related heart glycosides ouabain (1 microM) and digoxin (1 microM) are neither transported by bovine Oatp1a2 nor by human OATP1A2. We conclude that based on the tested substrates bovine Oatp1a2 shows functional homology to human OATP1A2.
Geyer
Joachim
J
Institute of Pharmacology and Toxicology, Justus-Liebig-University of Giessen, Frankfurter Str. 107, D-35392 Giessen, Germany.
Döring
Barbara
B
Failing
Klaus
K
Petzinger
Ernst
E
D000818:Animals; D001483:Base Sequence; D001692:Biological Transport; D002417:Cattle; D003001:Cloning, Molecular / Q000379:methods*; D018076:DNA, Complementary / Q000235:genetics / Q000302:isolation & purification; D004958:Estradiol / Q000031:analogs & derivatives* / Q000378:metabolism; D004970:Estrone / Q000031:analogs & derivatives* / Q000378:metabolism; D007700:Kinetics; D008565:Membrane Proteins; D008969:Molecular Sequence Data; D027361:Organic Anion Transporters / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism*; D016415:Sequence Alignment; D013656:Taurocholic Acid / Q000378:metabolism; D014018:Tissue Distribution
D018076:DNA, Complementary; D008565:Membrane Proteins; D027361:Organic Anion Transporters; C512988:SLCO1A2 protein, human; C025483:estradiol-17 beta-glucuronide; D004970:Estrone; D004958:Estradiol; D013656:Taurocholic Acid; C017296:estrone sulfate