Posttranslational modifications on protein kinase c isozymes. Effects of epinephrine and phorbol esters

Biochim Biophys Acta. 2008 May;1783(5):695-712. doi: 10.1016/j.bbamcr.2007.07.011. Epub 2007 Aug 22.

Abstract

The posttranslational modifications induced on PKC isozymes as result of their activation were investigated. Reciprocal immunoprecipitations followed by Western blot analysis demonstrated that all PKC isozymes expressed in rat hepatocytes are modified by tyrosine nitration and tyrosine phosphorylation in different ways upon exposure of cells to a direct PKC activator (TPA), or to an extracellular ligand known to activate PKC-dependent pathways (epinephrine). Our data demonstrate for the first time that all PKC isozymes are also dynamically modified by O-linked beta-N-acetylglucosamine (O-GlcNAc); the presence of this modification was confirmed in part by FT-ICR mass spectrometry analysis. Interestingly, the O-GlcNAc modified Ser or Thr were mapped at similar positions in several PKC isozymes. The biochemical meaning of these posttranslational modifications was investigated for PKC alpha and delta. It was found that the PKC phosphorylation status of both isozymes in tyrosine and serine residues seems to regulate directly the enzyme activity since catalytic inactivation correlate with dephosphorylation of Ser at the C-terminus autophosphorylation sites of each PKC isozyme, and with an increase in the level of tyrosine phosphorylation. Whereas none of the other posttranslational modifications showed per se a direct effect in PKC delta activity, increased tyrosine nitration and O-GlcNAc modifications correlate negatively with PKCalpha activity.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Acetylglucosamine / metabolism
  • Animals
  • Enzyme Activation
  • Epinephrine / pharmacology*
  • Hepatocytes / drug effects
  • Hepatocytes / enzymology
  • Isoenzymes / chemistry
  • Isoenzymes / metabolism
  • Male
  • Mass Spectrometry
  • Protein Kinase C / chemistry
  • Protein Kinase C / metabolism*
  • Protein Processing, Post-Translational*
  • Rats
  • Rats, Wistar
  • Serine / metabolism
  • Tetradecanoylphorbol Acetate / pharmacology*
  • Threonine / metabolism
  • Tyrosine / metabolism

Substances

  • Isoenzymes
  • Threonine
  • Tyrosine
  • Serine
  • Protein Kinase C
  • Tetradecanoylphorbol Acetate
  • Acetylglucosamine
  • Epinephrine