ENDOTHELIAL CELL HYPOXIA-ASSOCIATED PROTEINS

在体内，生物体及其驻留的组织和细胞经常 暴露在环境氧气减少的环境中。因此，对急性呼吸道感染的耐受性 缺氧，在某些情况下，对慢性缺氧的适应是关键的 存活并在不同的细胞中有所不同。一些自适应的 已经提出了允许发展耐缺氧的策略 和评估；其他人可能是未知的。我们最近发现了一种 耐低氧内皮细胞诱导的一组特定应激蛋白 细胞暴露在急性和慢性低氧环境中。这些蛋白质， 被称为缺氧相关蛋白(HAPS)的蛋白是由 环境低氧，与其他已描述的应激蛋白不同， 例如热休克或葡萄糖调节蛋白，而且很可能 在RNA水平上进行调控。为了表征内皮细胞的HAPS和 他们的监管更全面，我们会：1)研究医保计划本地化 在肺动脉内皮细胞内；2)研究 HAPS与其他应激蛋白之间的关系 低氧、热休克和缺糖的毒性作用；3)分离 HAPS蛋白直接测序及低氧条件下HAPS基因的克隆 刺激的肺动脉内皮细胞c DNA文库；4)使用 分离蛋白或由HAPS cDNA推导的蛋白序列 开发分子探针；5)使用抗体和cDNAs探针 研究HAPS的转录和翻译调控 培养的细胞和低氧组织，并确定其对 改变内皮细胞对缺氧反应的HAPS表达。这些 研究将提供对HAPS在生物学上的调节的洞察 和遗传水平，并可能为细胞内的缺氧和 纸巾。

In vivo, organisms and their resident tissues and cells are frequently exposed to decreases in ambient oxygen. Therefore, tolerance to acute hypoxia and, in some cases, adaptation to chronic hypoxia, is critical to survival and varies among different cells. Some of the adaptive strategies permitting development of hypoxia tolerance have been proposed and evaluated; others are likely unknown. We have recently identified a specific set of stress proteins induced in hypoxia tolerant endothelial cells during exposure to both acute and chronic hypoxia. These proteins, termed hypoxia associated proteins (HAPs), are specifically induced by environmental hypoxia, are distinct from other described stress proteins, such as heat shock or glucose-regulated proteins, and are likely regulated at the RNA level. To characterize endothelial cell HAPs and their regulation more fully, we will: 1) examine the localization of HAPs within pulmonary arterial endothelial cells; 2) investigate the relationship of HAPs and other stress proteins as protection against the toxic effects of hypoxia, heat shock, and glucose deprivation; 3) isolate HAPs by direct protein sequencing and by cloning HAPs cDNA from a hypoxia stimulated pulmonary arterial endothelial cell cDNA library; 4) use the isolated protein or the protein sequence deduced from HAPs cDNA to develop molecular probes; and 5) use antibody and cDNA probes to investigate transcriptional and translational regulation of HAPs in cultured cells and hypoxic tissues and to determine the effect of altering HAPs expression on endothelial cell response to hypoxia. These studies will provide insight into the regulation of HAPs at the biologic and genetic level and may provide markers for hypoxia in cells and tissues.