title: S-Nitrosation of β-catenin and p120 catenin: a novel regulatory mechanism in endothelial hyperpermeability.
abstract: BACKGROUND
  Endothelial adherens junction proteins constitute an important element in the control of microvascular permeability. Platelet-activating factor (PAF) increases permeability to macromolecules via translocation of endothelial nitric oxide synthase (eNOS) to cytosol and stimulation of eNOS-derived nitric oxide signaling cascade. The mechanisms by which nitric oxide signaling regulates permeability at adherens junctions are still incompletely understood.
  
  
  OBJECTIVE
  We explored the hypothesis that PAF stimulates hyperpermeability via S-nitrosation (SNO) of adherens junction proteins.
  
  
  RESULTS
  We measured PAF-stimulated SNO of β-catenin and p120-catenin (p120) in 3 cell lines: ECV-eNOSGFP, EAhy926 (derived from human umbilical vein), and postcapillary venular endothelial cells (derived from bovine heart endothelium) and in the mouse cremaster muscle in vivo. SNO correlated with diminished abundance of β-catenin and p120 at the adherens junction and with hyperpermeability. Tumor necrosis factor-α increased nitric oxide production and caused similar increase in SNO as PAF. To ascertain the importance of eNOS subcellular location in this process, we used ECV-304 cells transfected with cytosolic eNOS (GFPeNOSG2A) and plasma membrane eNOS (GFPeNOSCAAX). PAF induced SNO of β-catenin and p120 and significantly diminished association between these proteins in cells with cytosolic eNOS but not in cells wherein eNOS is anchored to the cell membrane. Inhibitors of nitric oxide production and of SNO blocked PAF-induced SNO and hyperpermeability, whereas inhibition of the cGMP pathway had no effect. Mass spectrometry analysis of purified p120 identified cysteine 579 as the main S-nitrosated residue in the region that putatively interacts with vascular endothelial-cadherin.
  
  
  CONCLUSIONS
  Our results demonstrate that agonist-induced SNO contributes to junctional membrane protein changes that enhance endothelial permeability.
authors: Instituto de Inmunología, Universidad Austral de Chile, Los Laureles s/n, 511-0566, Valdivia, Chile.
authors: Natalie
authors: N
authors: Marín
authors: Patricia
authors: P
authors: Zamorano
authors: Rodrigo
authors: R
authors: Carrasco
authors: Patricio
authors: P
authors: Mujica
authors: Francisco G
authors: FG
authors: González
authors: Claudia
authors: C
authors: Quezada
authors: Cynthia J
authors: CJ
authors: Meininger
authors: Mauricio P
authors: MP
authors: Boric
authors: Walter N
authors: WN
authors: Durán
authors: Fabiola A
authors: FA
authors: Sánchez
mesh_terms: D022005:Adherens Junctions / Q000378:metabolism*; D000595:Amino Acid Sequence; D000818:Animals; D002199:Capillary Permeability / Q000187:drug effects / Q000502:physiology*; D051177:Catenins / Q000235:genetics / Q000378:metabolism*; D002417:Cattle; D042783:Endothelial Cells / Q000378:metabolism*; D049452:Green Fluorescent Proteins / Q000235:genetics; D061307:Human Umbilical Vein Endothelial Cells; D006801:Humans; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D008822:Mice, Transgenic; D008969:Molecular Sequence Data; D018482:Muscle, Skeletal / Q000098:blood supply / Q000378:metabolism; D009569:Nitric Oxide / Q000378:metabolism; D052250:Nitric Oxide Synthase Type III / Q000235:genetics / Q000378:metabolism; D015538:Nitrosation / Q000502:physiology; D010982:Platelet-Derived Growth Factor / Q000494:pharmacology; D015398:Signal Transduction / Q000187:drug effects / Q000502:physiology*; D014699:Venules / Q000166:cytology; D051176:beta Catenin / Q000378:metabolism*; D000095002:Delta Catenin
chemical_list: C495270:CTNNB1 protein, human; C495267:CTNNB1 protein, mouse; D051177:Catenins; D010982:Platelet-Derived Growth Factor; D051176:beta Catenin; D049452:Green Fluorescent Proteins; D009569:Nitric Oxide; D052250:Nitric Oxide Synthase Type III; C496315:Nos3 protein, mouse; D000095002:Delta Catenin