p47phox associates with the cytoskeleton through cortactin in human vascular smooth muscle cells: role in NAD(P)H oxidase regulation by angiotensin II.
p47phox associates with the cytoskeleton through cortactin in human vascular smooth muscle cells: role in NAD(P)H oxidase regulation by angiotensin II.
复制标题
p47phox 通过人血管平滑肌细胞中的皮质素与细胞骨架结合：在血管紧张素 II 的 NAD(P)H 氧化酶调节中的作用。
DOI：
10.1161/01.atv.0000154141.66879.98
复制
发表时间：
2005
期刊：
影响因子：
--
通讯作者：
Schiffrin,EL
中科院分区：
文献类型：
--
作者：
Touyz,RM;Yao,G;Quinn,MT;Pagano,PJ;Schiffrin,EL
我们检验了p47 phox与肌动蛋白细胞骨架相关的假设，使NAD（P）H氧化酶的定点激活成为可能，并评估这些作用是否影响活性氧（ROS）的产生和血管紧张素II（Ang II）在来自人阻力和冠状动脉的血管平滑肌细胞（VSMC）中的信号传导。介导的O2生成，建立了在这种反应中对p47 phox的需求。免疫荧光共聚焦显微镜显示p47 phox在基础条件下的胞浆分布。在Ang II刺激后，p47 phox以线性方式重排，与F-actin共定位。免疫共沉淀研究证实了p47 phox和肌动蛋白之间的关联，并证明了与肌动蛋白结合蛋白corpine的相互作用。细胞骨架破坏与细胞松弛素阻止p47 phox：肌动蛋白的相互作用和衰减ROS的形成和p38 MAP激酶和Akt磷酸化的血管紧张素II。细胞内ROS的产生响应LY 83583（O2 generator）或外源性H2 O2和Ang II诱导的ERK 1/2激活被cytochalasin.Conclusions-The p47 phox：actin相互作用，通过coronin，在Ang II介导的功能活性NAD（P）H氧化酶的定点组装，ROS的产生和氧化还原敏感的p38 MAP激酶和Akt的激活中起着重要作用，但不是ERK 1/2。这些发现表明，在人VSMCs中，完整的肌动蛋白细胞骨架在NAD（P）H氧化酶调节和Ang II的氧化还原信号传导中的重要性。
Objective—We tested the hypothesis that p47phox associates with the actin cytoskeleton, enabling site-directed activation of NAD(P)H oxidase, and assessed whether these actions influence reactive oxygen species (ROS) generation and signaling by angiotensin II (Ang II) in vascular smooth muscle cells (VSMCs) from human resistance and coronary arteries.Methods and Results—Electroporation of anti-p47phox antibody into VSMCs abrogated Ang II-mediated O2generation, establishing the requirement for p47phox in this response. Immunfluorescence confocal microscopy demonstrated a cytosolic distribution of p47phox in basal conditions. After Ang II stimulation, p47phox rearranged in a linear fashion, colocalizing with F-actin. Co-immunoprecipitation studies confirmed an association between p47phox and actin and demonstrated an interaction with the actin-binding protein cortactin. Cytoskeletal disruption with cytochalasin prevented p47phox:actin interaction and attenuated ROS formation and p38MAP kinase and Akt phosphorylation by Ang II. Intracellular ROS generation in response to LY83583 (O2generator) or exogenous H2O2and Ang II-induced ERK1/2 activation were unaltered by cytochalasin.Conclusions—The p47phox:actin interaction, through cortactin, plays an important role in Ang II-mediated site-directed assembly of functionally active NAD(P)H oxidase, ROS generation, and activation of redox-sensitive p38MAP kinase and Akt, but not ERK1/2. These findings demonstrate the importance of an intact actin-cytoskeleton in NAD(P)H oxidase regulation and redox signaling by Ang II in human VSMCs.