Differential modulation of a polymorphism in the COOH terminus of the α-subunit of the human epithelial sodium channel by protein kinase Cδ

Wusheng Yan, Laurence Suaud, Thomas R. Kleyman, Ronald C. Rubenstein

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Abstract

The A663T polymorphism of the α-subunit of the human epithelial sodium channel (hENaC) increases the functional and surface expression of αβγ-hENaC in Xenopus laevis oocytes. The context of this residue in the COOH terminus of α-hENaC is important for this effect, as a homologous change in murine ENaC (mENaC), A692T, does not alter functional and surface expression of mENaC. Query of a phosphoprotein database suggested that the α-T663 residue might be phosphorylated by PKCδ. General inhibition of PKC with calphostin C decreased the functional and surface expression of αT663-hENaC and not αA663-hENaC, and was without effect on αA692-mENaC, αT692-mENaC, and a chimeric m(1-678)/h(650-669)αT663, mβγ-ENaC. These data suggest that residues outside of the α-hENaC COOH terminus are important for modulation of αT663-hENaC trafficking by PKC. In contrast, expression of PKCδ decreased the functional and surface expression of αT663-hENaC and the functional expression of m(1-678)/h(650-669)αT663, mβγ-ENaC, and was without effect on αA663-hENaC, αA692-mENaC, or αT692-mENaC. PKCδ did not phosphorylate the COOH terminus of either αT663-hENaC or αA663-hENaC in vitro, suggesting that it acts indirectly to regulate hENaC trafficking. αT663-hENaC was retrieved from the oocyte membrane more slowly than αA663-hENaC, and calphostin C increased the rate of αT663-hENaC removal from the oocyte membrane to a rate similar to that of αA663-hENaC. In contrast, PKCδ did not alter the rate of removal of αT663-hENaC from the oocyte membrane, suggesting that PKCδ altered rates of αT663-hENaC biosynthesis and/or delivery to the plasma membrane. These data are consistent with PKC isoform-specific effects on the intracellular trafficking of αT663- vs. αA663-hENaC.

Original languageEnglish
Pages (from-to)F279-F288
JournalAmerican Journal of Physiology - Renal Physiology
Volume290
Issue number2
DOIs
StatePublished - Feb 2006

Keywords

  • Oocyte
  • Phosphorylation
  • Trafficking

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