Abstract
We characterized the interactions of various compounds with OAT-K1 and OAT-K2, kidney-specific organic anion transporters. By using Madin-Darby canine kidney cells stably transfected with OAT-K1 or OAT-K2 cDNA, the antitumor drug methotrexate, the mycotoxin ochratoxin A, endogenous organic anions (thyroid hormones, taurocholic acid, and conjugated steroids), and the antiretroviral drug zidovudine were shown to be substrates for these transporters. Although the apparent Michaelis constant (Km) values of methotrexate for OAT-K1 and OAT-K2 were 2.1 and 1.8 μM, respectively, 2.5 mM methotrexate inhibited only 20% of the 125I-thyroid hormones uptake via these transporters. In addition, 100 μM methotrexate did not have any effect on [3H]zidovudine uptake via OAT-K1 or OAT-K2. Similarly, several substrates caused little or no mutual inhibition at concentrations much higher than theirKm values for these transporters. Moreover, intracellular methotrexate trans-stimulated the OAT-K1- and OAT-K2-mediated uptake of [3H]folic acid, but not that of other compounds. Organic anion-transporting polypeptide 2 (oatp2), a liver-type homolog of OAT-K1 and OAT-K2, showed similar events. The inhibition constant values of triiodothyronine and taurocholic acid for [3H]digoxin uptake in oatp2-expressing oocytes resulted in 50.4 and 1.48 mM, respectively, which were about 9- and 40-fold higher than theirKm values for oatp2, respectively. These findings suggested that several substrates interact with these transporters at different amino acid residue(s). Taken together, these observations suggested that OAT-K1 and OAT-K2 could serve as multispecific transporters, mediating transport of a wide variety of endogenous substances, xenobiotics, and their metabolites in the kidney, presumably via several interaction sites in their molecules.
Footnotes
-
This work was supported by a grant-in-aid for scientific research from Ministry of Education, Science, and Culture of Japan.
- Abbreviations:
- OAT
- organic anion transporter
- MDCK
- Madin-Darby canine kidney
- oatp
- organic anion-transporting polypeptide
- DHEA
- dehydroepiandrosterone
- rOAT1
- rat organic anion transporter 1
- Received April 26, 2001.
- Accepted June 19, 2001.
- The American Society for Pharmacology and Experimental Therapeutics
JPET articles become freely available 12 months after publication, and remain freely available for 5 years.Non-open access articles that fall outside this five year window are available only to institutional subscribers and current ASPET members, or through the article purchase feature at the bottom of the page.
|