WNK4 enhances the degradation of NCC through a sortilin-mediated lysosomal pathway

J Am Soc Nephrol. 2010 Jan;21(1):82-92. doi: 10.1681/ASN.2008121275. Epub 2009 Oct 29.

Abstract

WNK kinase is a serine/threonine kinase that plays an important role in electrolyte homeostasis. WNK4 significantly inhibits the surface expression of the sodium chloride co-transporter (NCC) by enhancing the degradation of NCC through a lysosomal pathway, but the mechanisms underlying this trafficking are unknown. Here, we investigated the effect of the lysosomal targeting receptor sortilin on NCC expression and degradation. In Cos-7 cells, we observed that the presence of WNK4 reduced the steady-state amount of NCC by approximately half. Co-transfection with truncated sortilin (a dominant negative mutant) prevented this WNK4-induced reduction in NCC. NCC immunoprecipitated with both wild-type sortilin and, to a lesser extent, truncated sortilin. Immunostaining revealed that WNK4 increased the co-localization of NCC with the lysosomal marker cathepsin D, and NCC co-localized with wild-type sortilin, truncated sortilin, and WNK4 in the perinuclear region. These findings suggest that WNK4 promotes NCC targeting to the lysosome for degradation via a mechanism involving sortilin.

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • Adaptor Proteins, Vesicular Transport / genetics
  • Adaptor Proteins, Vesicular Transport / metabolism*
  • Animals
  • COS Cells
  • Cathepsin D / metabolism
  • Cell Line
  • Chlorocebus aethiops
  • Humans
  • Kidney Tubules, Distal / cytology
  • Kidney Tubules, Distal / metabolism*
  • Lysosomes / metabolism*
  • Mutation
  • Protein Serine-Threonine Kinases / metabolism*
  • Protein Transport / physiology
  • Signal Transduction / physiology*
  • Sodium Chloride Symporters / metabolism*
  • Transfection

Substances

  • Adaptor Proteins, Vesicular Transport
  • Sodium Chloride Symporters
  • Protein Serine-Threonine Kinases
  • WNK4 protein, human
  • Cathepsin D
  • sortilin