WNK1 phosphorylates synaptotagmin 2 and modulates its membrane binding

Mol Cell. 2004 Sep 10;15(5):741-51. doi: 10.1016/j.molcel.2004.07.018.

Abstract

WNK (with no lysine [K]) protein kinases were named for their unique active site organization. Mutations in WNK1 and WNK4 cause a familial form of hypertension by undefined mechanisms. Here, we report that WNK1 selectively binds to and phosphorylates synaptotagmin 2 (Syt2) within its calcium binding C2 domains. Endogenous WNK1 and Syt2 coimmunoprecipitate and colocalize on a subset of secretory granules in INS-1 cells. Phosphorylation by WNK1 increases the amount of Ca2+ required for Syt2 binding to phospholipid vesicles; mutation of threonine 202, a WNK1 phosphorylation site, partially prevents this change. These findings suggest that phosphorylation of Syts by WNK1 can regulate Ca2+ sensing and the subsequent Ca2+-dependent interactions mediated by Syt C2 domains. These findings provide a biochemical mechanism that could lead to the retention or insertion of proteins in the plasma membrane. Interruption of this regulatory pathway may disturb membrane events that regulate ion balance.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Animals
  • Binding Sites / physiology
  • Calcium Signaling / genetics
  • Calcium-Binding Proteins / metabolism
  • Cell Line
  • Cell Membrane / metabolism*
  • Humans
  • Intracellular Signaling Peptides and Proteins
  • Minor Histocompatibility Antigens
  • Mutation / genetics
  • Nerve Tissue Proteins / metabolism*
  • Phosphorylation
  • Protein Binding / physiology*
  • Protein Serine-Threonine Kinases / genetics
  • Protein Serine-Threonine Kinases / metabolism*
  • Protein Structure, Tertiary / physiology
  • Secretory Vesicles / metabolism
  • Synaptotagmin II
  • Threonine / metabolism
  • Two-Hybrid System Techniques
  • WNK Lysine-Deficient Protein Kinase 1
  • Water-Electrolyte Balance

Substances

  • Calcium-Binding Proteins
  • Intracellular Signaling Peptides and Proteins
  • Minor Histocompatibility Antigens
  • Nerve Tissue Proteins
  • SYT2 protein, human
  • Synaptotagmin II
  • Threonine
  • Protein Serine-Threonine Kinases
  • WNK Lysine-Deficient Protein Kinase 1
  • WNK1 protein, human