• DOI:10.1152/AJPRENAL.00161.2003
• Corpus ID: 907020

Regulation of sodium-dicarboxylate cotransporter-3 from winter flounder kidney by protein kinase C.

@article{Hagos2004RegulationOS,
  title={Regulation of sodium-dicarboxylate cotransporter-3 from winter flounder kidney by protein kinase C.},
  author={Yohannes Hagos and Birgitta Christina Burckhardt and Alexander Larsen and Christian Mathys and Tobias Gronow and Andrew Bahn Bahn and Natascha A. Wolff and Gerhard Burckhardt and J{\"u}rgen Steffgen},
  journal={American journal of physiology. Renal physiology},
  year={2004},
  volume={286 1},
  pages={
          F86-93
        }
}

• Y. Hagos, B. Burckhardt, +6 authors J. Steffgen
• Published 2004
• Biology
• American journal of physiology. Renal physiology

The sodium dicarboxylate cotransporter located at the basolateral side supplies renal proximal tubule cells with Krebs cycle intermediates and maintains the driving force for the exchange of organic anions like PAH against alpha-ketoglutarate through the organic anion transporter-1. Recently, we cloned sodium dicarboxylate cotransporter-3 from winter flounder kidney (fNaDC-3). To understand the regulation of fNaDC-3, we preincubated fNaDC-3-expressing oocytes with PMA, a PKC activator. PMA dose and time dependently inhibited fNaDC-3-mediated succinate uptake. Simultaneous preincubation of fNaDC-3-expressing oocytes with 50 nM PMA and either staurosporine or RO 31-8220 for 30 min attenuated PKC-mediated inhibition of succinate uptake. Site-directed mutagenesis of the five putative PKC sites (S7, T167, S174, T188, and S396) resulted in no change in PKC-mediated inhibition of the transporter. In electrophysiological studies performed at -60 mV, the K0.5 for succinate was not significantly affected (56 +/- 13 vs. 42 +/- 19 microM), but DeltaImax was reduced from -139 +/- 49 to -20 +/- 8 nA by PMA (50 nM, 30 min). Immunofluorescence analysis of fNaDC-3-expressing oocytes revealed that PMA leads to an endocytosis of fNaDC-3 protein. In conclusion, fNaDC-3 expressed in oocytes is downregulated by PMA through endocytosis. PKC consensus sites appear not to be important for this process.