p47 binding partners in endothelial cell function

描述(由申请人提供)：缺血性和炎症性疾病 靶向血管内皮细胞，并以升高的水平为标志 细胞因子、强烈的氧化应激和血管通透性增加。这个 血管这三个特征之间关系的确切性质 然而，疾病的定义不明确，阻碍了治疗这些常见疾病的努力 综合症。 直到最近，人们才意识到氧化剂参与了生理学 信号传递过程，尤其是由肿瘤坏死因子-α启动的信号传递过程，尽管 这种氧化剂的来源和目标尚不清楚。在简单的体外系统中， 氧化剂的转瞬即逝的性质使得它们的影响具有高度的部位特异性， 使得它们的起源位置通常决定了它们的分子靶标。 最近的研究还表明，信号的特异性，特别是在 MAP-Kinase系统，也依赖于信号元件的适当共定位。 我们的初步数据表明，吞噬细胞氧化酶样复合体在 肿瘤坏死因子-α诱导MAP激酶c-jun氨基末端激酶的激活 其活性形式与人内皮细胞中的细胞骨架有关 细胞。我们克隆了一种重要的信号受体蛋白p47Phox。 从人类内皮细胞库中提取的酶，发现 P47Phox蛋白也定位于细胞骨架。此外，我们已经取得了 并筛选了p47Phox结合伙伴的HUVEC GAL4文库，并具有 恢复了赠款正文中详细说明的几个重要基因。 我们假设炎症信号通过细胞因子和生长因子在 血管细胞涉及吞噬细胞特定部位氧化剂的产生 氧化物型复合体，导致JNK和NF-KB下游活化。我们 进一步提出该酶与细胞骨架相关 元素通过与信号蛋白的特定相互作用。 我们的总体目标将是研究p47Phox的性质。 与三个潜在结合靶点的相互作用，并探索 这些互动的功能意义通过检查上游和 下行信令事件。通过更详细地了解 细胞因子诱导的促炎事件，将出现更多机会 设计针对缺血和炎症血管的具体干预措施 疾病。

DESCRIPTION (provided by the applicant): Ischemic and inflammatory diseases target the vascular endothelium and are marked by increased levels of cytokines, intense oxidative stress, and increased vascular permeability. The precise nature of the relationship between these three features of vascular diseases is ill defined, however, impeding efforts to treat these common syndromes. Oxidants have only recently been appreciated as participating in physiologic signaling processes, particularly those initiated by TNF-alpha, although the source and targets of such oxidants are not known. In simple in vitro systems, the evanescent nature of oxidants renders their effects highly site-specific, such that their site of origin generally dictates their molecular target. Recent studies have also revealed that signal specificity, particularly in the MAP Kinase system, also depends on proper colocalization of signaling elements. Our preliminary data implicate a phagocyte oxidase-like complex in the TNF-alpha induced activation of c-Jun amino terminal kinase (JNK), a MAP kinase whose active form is associated with the cytoskeleton, in human endothelial cells. We have cloned p47phox, an important signal-receiving adapter protein for the oxidase, from a human endothelial cell library, and found that the p47phox protein is also localized to the cytoskeleton. Further, we have made and screened a HUVEC GAL4 library for binding partners of p47phox, and have recovered several important genes detailed in the body of the grant. We hypothesize that inflammatory signaling by cytokines and growth factors in vascular cells involves site-specific production of oxidants by a phagocyte oxidase-like complex, which leads to downstream activation of JNK and NF-KB. We further propose that the oxidase is tethered to cytoskeleton-associated elements through specific interactions with signaling proteins. Our overall objectives will be to investigate the nature of p47phox interactions with three potential binding targets, and to explore the functional significance of these interactions by examining upstream and downstream signaling events. Through a more detailed understanding of cytokine-induced proinflammatory events, more opportunities will arise for designing specific interventions against ischemic and inflammatory vascular diseases.