American Journal of Physiology-Gastrointestinal and Liver Physiology · 2000 · 620 citations · 38 references
Life SciencesBiochemistryMolecular BiologyCellular BiochemistryMembrane BiochemistryProtein BiochemistryHepatic Drug AbsorptionProtein TargetingBasolateral MembraneBasolateral TransportersHek293 Cell ExpressionHepatic TransportersLateral Membrane DomainOatp-mediated UptakeOrganic Anion Transporting Polypeptide-mediated TransportTransporter Expression In Hepatocytes
We cloned and expressed a new organic anion transporting polypeptide (OATP), termed human OATP2, (OATP-C, LST-1; symbol SLC21A6), involved in the uptake of various lipophilic anions into human liver. The cDNA encoding OATP2 comprised 2073 base pairs, corresponding to a protein of 691 amino acids, which were 44% identical to the known human OATP. An antibody directed against the carboxy terminus localized OATP2 to the basolateral membrane of human hepatocytes. Northern blot analysis indicated a strong expression of OATP2 only in human liver. Transport mediated by recombinant OATP2 and its localization were studied in stably transfected Madin-Darby canine kidney strain II (MDCKII) and HEK293 cells. Confocal microscopy localized recombinant OATP2 protein to the lateral membrane of MDCKII cells. Substrates included 17beta-glucuronosyl estradiol, monoglucuronosyl bilirubin, dehydroepiandrosterone sulfate, and cholyltaurine. 17beta-Glucuronosyl estradiol was a preferred substrate, with a Michaelis-Menten constant value of 8.2 microM; its uptake was Na(+) independent and was inhibited by sulfobromophthalein, with a inhibition constant value of 44 nM. Our results indicate that OATP2 is important for the uptake of organic anions, including bilirubin conjugates and sulfobromophthalein, in human liver.
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A comprehensive set of sequence analysis programs for the VAX
John Devereux, Paul Haeberli, Oliver Smithies · Nucleic Acids Research · 1984 · 14.4K citations · Full text
The Determination of Enzyme Dissociation Constants
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