Inhibition of PKCalpha and rhoA translocation in differentiated smooth muscle by a caveolin scaffolding domain peptide

Exp Cell Res. 2000 Jul 10;258(1):72-81. doi: 10.1006/excr.2000.4891.

Abstract

Receptor-coupled contraction of smooth muscle involves recruitment to the plasma membrane of downstream effector molecules PKCalpha and rhoA but the mechanism of this signal integration is unclear. Caveolins, the principal structural proteins of caveolar plasma membrane invaginations, have been implicated in the organization and regulation of many signal transducing molecules. Thus, using laser scanning confocal immunofluorescent microscopy, we tested the hypothesis that caveolin is involved in smooth muscle signaling by investigating caveolin isoform expression and localization, together with the effect of a peptide inhibitor of caveolin function, in intact differentiated smooth muscle cells. All three main caveolin isoforms were identified in uterine, stomach, and ileal smooth muscles and assumed a predominantly plasma membranous localization in myometrial cells. Cytoplasmic introduction of a peptide corresponding to the caveolin-1 scaffolding domain-an essential region for caveolin interaction with signaling molecules--significantly inhibited agonist-induced translocation of both PKCalpha and rhoA. Translocation was unimpaired by a scrambled peptide and was unaltered in sham-treated cells. The membranous localization of caveolins, and direct inhibition of receptor-coupled PKCalpha and rhoA translocation by the caveolin-1 scaffolding domain, supports the concept that caveolins can regulate the integration of extracellular contractile stimuli and downstream intracellular effectors in smooth muscle.

Publication types

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

MeSH terms

  • Amino Acid Sequence
  • Animals
  • Caveolin 1
  • Caveolin 2
  • Caveolins*
  • Cell Differentiation
  • Female
  • Isoenzymes / antagonists & inhibitors
  • Isoenzymes / metabolism*
  • Membrane Proteins / chemistry
  • Membrane Proteins / physiology*
  • Molecular Sequence Data
  • Muscle, Smooth / cytology*
  • Muscle, Smooth / drug effects
  • Muscle, Smooth / physiology*
  • Peptide Fragments / pharmacology*
  • Pregnancy
  • Protein Kinase C / antagonists & inhibitors
  • Protein Kinase C / metabolism*
  • Protein Kinase C-alpha
  • Rats
  • Rats, Sprague-Dawley
  • Uterus / cytology
  • Uterus / physiology
  • rhoA GTP-Binding Protein / antagonists & inhibitors
  • rhoA GTP-Binding Protein / metabolism*

Substances

  • Cav1 protein, rat
  • Caveolin 1
  • Caveolin 2
  • Caveolins
  • Isoenzymes
  • Membrane Proteins
  • Peptide Fragments
  • Protein Kinase C
  • Protein Kinase C-alpha
  • rhoA GTP-Binding Protein