Regulation by protein kinase C of organic anion transport driven by rat organic anion transporter 3 (rOAT3).
The organic anion transporter 3 (rOAT3) is a multispecific OAT localized at the basolateral membrane of the proximal tubule. The purpose of this study was to elucidate the role of protein kinase C (PKC) in the regulation of organic anion transport driven by rOAT3 and its mechanism of action. For this purpose, we established and utilized cells derived from the second segment of proximal tubule from mice stably expressing rOAT3 (S2 rOAT3). Phorbol 12-myristate 13-acetate (PMA), a PKC stimulator, attenuated the cellular uptake of estrone sulfate (ES), a prototype organic anion for rOAT3, in a dose- and time-dependent manner. PMA treatment resulted in a decrease in the Vmax, but not the Km of uptake of ES in S2 rOAT3. Treatment of S2 rOAT3 with other PKC stimulators or diacylglycerols also inhibited the uptake of ES, whereas that with an inactive phorbol ester did not. Chelerythrine chloride, a PKC inhibitor, reversed the PMA-induced decrease in uptake of ES in S2 rOAT3. These results suggest that PKC activation downregulates rOAT3-mediated organic anion transport. This down-regulation may be due to the inhibition of translocation or internalization of the rOAT3 protein, resulting in the decrease in the Vmax of rOAT3-mediated organic anion transport.
Takeda
M
M
Department of Pharmacology and Toxicology, Kyorin University School of Medicine, Tokyo, Japan.
Sekine
T
T
Endou
H
H
D000818:Animals; D000838:Anions / Q000493:pharmacokinetics; D001693:Biological Transport, Active; D002352:Carrier Proteins / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism*; D002478:Cells, Cultured; D004305:Dose-Response Relationship, Drug; D004789:Enzyme Activation; D004970:Estrone / Q000031:analogs & derivatives / Q000493:pharmacokinetics*; D007687:Kidney Tubules, Proximal / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D051379:Mice; D008822:Mice, Transgenic; D029362:Organic Anion Transporters, Sodium-Independent; D011493:Protein Kinase C / Q000378:metabolism / Q000502:physiology*; D013379:Substrate Specificity; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology; D014162:Transfection
D000838:Anions; D002352:Carrier Proteins; D029362:Organic Anion Transporters, Sodium-Independent; C114338:organic anion transport protein 3; D004970:Estrone; D011493:Protein Kinase C; D013755:Tetradecanoylphorbol Acetate; C017296:estrone sulfate