Rab proteins regulate epithelial sodium channel activity in colonic epithelial HT-29 cells

Sunil Saxena, Madhurima Singh, Kathrin Engisch, Mitsunori Fukuda, Simarna Kaur

Research output: Contribution to journalArticlepeer-review

30 Citations (Scopus)

Abstract

ENaC, the sodium-selective amiloride-sensitive epithelial channel, mediates electrogenic sodium re-absorption in tight epithelia and is deeply associated with human hypertension. The ENaC expression at plasma membrane requires the regulated transport, processing, and macromolecular assembly in a defined and highly compartmentalized manner. Ras-related Rab GTPases regulate intracellular trafficking during endocytosis, regulated exocytosis, and secretion. To evaluate the role of these proteins in regulating amiloride-sensitive sodium channel activity, multiple Rab isoforms 3, 5, 6, and Rab27a were expressed in HT-29 cells. Rab3 and Rab27a inhibited ENaC currents, while the expression of other Rab isoforms failed to elicit any statistically significant effect on amiloride-sensitive currents. The immunoprecipitation experiments suggest protein-protein interaction of Rab3 and Rab27a with epithelial sodium channel. Biotinylation studies revealed that modulation of ENaC function is due to the reduced apical expression of channel proteins. Study also indicates that Rabs do not appear to affect the steady-state level of total cellular ENaC. Alternatively, introduction of isoform-specific small inhibitory RNA (SiRNA) reversed the Rab-dependent inhibition of amiloride-sensitive currents. These observations point to the involvement of multiple Rab proteins in ENaC transport through intracellular routes like exocytosis, recycling from ER to plasma membrane or degradation and thus serve as potential target for human hypertension.

Original languageEnglish
Pages (from-to)1219-1223
Number of pages5
JournalBiochemical and biophysical research communications
Volume337
Issue number4
DOIs
Publication statusPublished - 2005 Dec 2
Externally publishedYes

Keywords

  • Biotinylation
  • Colonic epithelial cells
  • ENaC
  • HT-29 cells
  • Protein-protein interaction
  • Rab27a
  • Rab3
  • Rab6
  • Regulation
  • Trafficking

ASJC Scopus subject areas

  • Biophysics
  • Biochemistry
  • Molecular Biology
  • Cell Biology

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