Modulation of Rac1 activity by ADMA/DDAH regulates pulmonary endothelial barrier function
Standard
Modulation of Rac1 activity by ADMA/DDAH regulates pulmonary endothelial barrier function. / Wojciak-Stothard, Beata; Torondel, Belen; Zhao, Lan; Renné, Thomas; Leiper, James M.
In: MOL BIOL CELL, Vol. 20, No. 1, 01.01.2009, p. 33-42.Research output: SCORING: Contribution to journal › SCORING: Journal article › Research › peer-review
Harvard
APA
Vancouver
Bibtex
}
RIS
TY - JOUR
T1 - Modulation of Rac1 activity by ADMA/DDAH regulates pulmonary endothelial barrier function
AU - Wojciak-Stothard, Beata
AU - Torondel, Belen
AU - Zhao, Lan
AU - Renné, Thomas
AU - Leiper, James M
PY - 2009/1/1
Y1 - 2009/1/1
N2 - Endogenously produced nitric oxide synthase inhibitor, asymmetric methylarginine (ADMA) is associated with vascular dysfunction and endothelial leakage. We studied the role of ADMA, and the enzymes metabolizing it, dimethylarginine dimethylaminohydrolases (DDAH) in the regulation of endothelial barrier function in pulmonary macrovascular and microvascular cells in vitro and in lungs of genetically modified heterozygous DDAHI knockout mice in vivo. We show that ADMA increases pulmonary endothelial permeability in vitro and in in vivo and that this effect is mediated by nitric oxide (NO) acting via protein kinase G (PKG) and independent of reactive oxygen species formation. ADMA-induced remodeling of actin cytoskeleton and intercellular adherens junctions results from a decrease in PKG-mediated phosphorylation of vasodilator-stimulated phosphoprotein (VASP) and a subsequent down-regulation of Rac1 activity. The effects of ADMA on endothelial permeability, Rac1 activation and VASP phosphorylation are prevented by overexpression of active DDAHI and DDAHII, whereas inactive DDAH mutants have no effect. These findings demonstrate for the first time that ADMA metabolism critically determines pulmonary endothelial barrier function by modulating Rac1-mediated remodeling of the actin cytoskeleton and intercellular junctions.
AB - Endogenously produced nitric oxide synthase inhibitor, asymmetric methylarginine (ADMA) is associated with vascular dysfunction and endothelial leakage. We studied the role of ADMA, and the enzymes metabolizing it, dimethylarginine dimethylaminohydrolases (DDAH) in the regulation of endothelial barrier function in pulmonary macrovascular and microvascular cells in vitro and in lungs of genetically modified heterozygous DDAHI knockout mice in vivo. We show that ADMA increases pulmonary endothelial permeability in vitro and in in vivo and that this effect is mediated by nitric oxide (NO) acting via protein kinase G (PKG) and independent of reactive oxygen species formation. ADMA-induced remodeling of actin cytoskeleton and intercellular adherens junctions results from a decrease in PKG-mediated phosphorylation of vasodilator-stimulated phosphoprotein (VASP) and a subsequent down-regulation of Rac1 activity. The effects of ADMA on endothelial permeability, Rac1 activation and VASP phosphorylation are prevented by overexpression of active DDAHI and DDAHII, whereas inactive DDAH mutants have no effect. These findings demonstrate for the first time that ADMA metabolism critically determines pulmonary endothelial barrier function by modulating Rac1-mediated remodeling of the actin cytoskeleton and intercellular junctions.
KW - Adherens Junctions
KW - Amidohydrolases
KW - Animals
KW - Antigens, CD
KW - Arginine
KW - Cadherins
KW - Cell Adhesion Molecules
KW - Cells, Cultured
KW - Cyclic GMP-Dependent Protein Kinases
KW - Cytoskeleton
KW - Endothelium
KW - Enzyme Activation
KW - Enzyme Inhibitors
KW - Isoenzymes
KW - Lung
KW - Mice
KW - Mice, Knockout
KW - Microfilament Proteins
KW - Nitric Oxide
KW - Nitric Oxide Synthase
KW - Permeability
KW - Phosphoproteins
KW - Reactive Oxygen Species
KW - Swine
KW - cdc42 GTP-Binding Protein
KW - rac1 GTP-Binding Protein
KW - rhoA GTP-Binding Protein
U2 - 10.1091/mbc.E08-04-0395
DO - 10.1091/mbc.E08-04-0395
M3 - SCORING: Journal article
C2 - 18923147
VL - 20
SP - 33
EP - 42
JO - MOL BIOL CELL
JF - MOL BIOL CELL
SN - 1059-1524
IS - 1
ER -