TNF-α potentiates protein-tyrosine nitration through activation of NADPH oxidase and eNOS localized in membrane rafts and caveolae of bovine aortic endothelial cells

被引:89
作者
Yang, Baohua
Rizzo, Victor
机构
[1] Temple Univ, Sch Med, Cardiovasc Res Ctr, Philadelphia, PA 19140 USA
[2] Temple Univ, Sch Med, Dept Anat & Cell Biol, Philadelphia, PA 19140 USA
来源
AMERICAN JOURNAL OF PHYSIOLOGY-HEART AND CIRCULATORY PHYSIOLOGY | 2007年 / 292卷 / 02期
关键词
lipid rafts; endothelial nitric oxide synthase; reactive oxygen species;
D O I
10.1152/ajpheart.00758.2006
中图分类号
R5 [内科学];
学科分类号
1002 ; 100201 ;
摘要
A major source of reactive oxygen species (ROS) in endothelial cells is the NADPH oxidase enzyme complex. The selective distributions of any enzyme within cells have important implications in regulating enzyme effectiveness through facilitation of access to local substrates and/or product targets. Because membrane rafts provide a spatially preferable environment for a variety of enzyme systems, we sought to determine whether NADPH oxidase is present and functional in this plasma membrane compartment in endothelial cells. We found that, in resting endothelial cells, NADPH oxidase subunits were preassembled and the enzyme functional in membrane rafts, specifically in caveolae. Stimulation with TNF-alpha induced additional recruitment of the p47(phox) regulatory subunit to raft-localized NADPH oxidase and enhanced ROS production within raft domains. TNF-alpha also induced nitric oxide production through activation of endothelial nitric oxide synthase (eNOS) present in the same membrane compartment. The dual activation of superoxide and nitric oxide-generating systems provided a spatially favorable environment for nitration of tyrosine-containing proteins localized to rafts. Perturbation of membrane raft structural integrity with cholesterol-sequestering compounds caused the delocalization of NADPH oxidase subunits and eNOS from the rafts and inhibited TNF-alpha-induced ROS production and protein tyrosine nitration. Together, these data provide evidence that membrane rafts and caveolae play a role in the spatial regulation of NADPH oxidase and subsequent ROS/reactive nitrogen species in endothelial cells.
引用
收藏
页码:H954 / H962
页数:9
相关论文
共 65 条
  • [11] Homologs of gp91phox:: cloning and tissue expression of Nox3, Nox4, and Nox5
    Cheng, GJ
    Cao, ZH
    Xu, XX
    Van Meir, EG
    Lambeth, JD
    [J]. GENE, 2001, 269 (1-2) : 131 - 140
  • [12] Caveolae participate in tumor necrosis factor receptor 1 signaling and internalization in a human endothelial cell line
    D'Alessio, A
    Al-Lamki, RS
    Bradley, JR
    Pober, JS
    [J]. AMERICAN JOURNAL OF PATHOLOGY, 2005, 166 (04) : 1273 - 1282
  • [13] Endothelin-1 enhances oxidative stress, cell proliferation and reduces apoptosis in human umbilical vein endothelial cells:: role of ETB receptor, NADPH oxidase and caveolin-1
    Dong, F
    Zhang, XC
    Wold, LE
    Ren, Q
    Zhang, ZJ
    Ren, J
    [J]. BRITISH JOURNAL OF PHARMACOLOGY, 2005, 145 (03) : 323 - 333
  • [14] Free radicals in the physiological control of cell function
    Dröge, W
    [J]. PHYSIOLOGICAL REVIEWS, 2002, 82 (01) : 47 - 95
  • [15] Depletion of plasma membrane cholesterol dampens hydrostatic pressure and shear stress-induced mechanotransduction pathways in osteoblast cultures
    Ferraro, JT
    Daneshmand, M
    Bizios, R
    Rizzo, V
    [J]. AMERICAN JOURNAL OF PHYSIOLOGY-CELL PHYSIOLOGY, 2004, 286 (04): : C831 - C839
  • [16] Cholesterol and caveolae: structural and functional relationships
    Fielding, CJ
    Fielding, PE
    [J]. BIOCHIMICA ET BIOPHYSICA ACTA-MOLECULAR AND CELL BIOLOGY OF LIPIDS, 2000, 1529 (1-3): : 210 - 222
  • [17] Phosphorylation of Thr495 regulates Ca2+/calmodulin-dependent endothelial nitric oxide synthase activity
    Fleming, I
    Fisslthaler, B
    Dimmeler, S
    Kemp, BE
    Busse, R
    [J]. CIRCULATION RESEARCH, 2001, 88 (11) : E68 - E75
  • [18] PKCζ regulates TNF-α-induced activation of NADPH oxidase in endothelial cells
    Frey, RS
    Rahman, A
    Kefer, JC
    Minshall, RD
    Malik, AB
    [J]. CIRCULATION RESEARCH, 2002, 90 (09) : 1012 - 1019
  • [19] Src kinase activates endothelial nitric-oxide synthase by phosphorylating Tyr-83
    Fulton, D
    Church, JE
    Ruan, L
    Li, CY
    Sood, SG
    Kemp, BE
    Jennings, IG
    Venema, RC
    [J]. JOURNAL OF BIOLOGICAL CHEMISTRY, 2005, 280 (43) : 35943 - 35952
  • [20] NAD(P)H oxidase mediates the endothelial barrier dysfunction induced by TNF-α
    Gertzberg, N
    Neumann, P
    Rizzo, V
    Johnson, A
    [J]. AMERICAN JOURNAL OF PHYSIOLOGY-LUNG CELLULAR AND MOLECULAR PHYSIOLOGY, 2004, 286 (01) : L37 - L48