ENDOTHELIAL CELL HYPOXIA-ASSOCIATED PROTEINS

内皮细胞缺氧相关蛋白

• 批准号: 2222221
• 负责人: HARRISON W FARBER
• 金额: $ 28.73万
• 依托单位: BOSTON UNIVERSITY MEDICAL CAMPUS
• 依托单位国家: 美国
• 财政年份: 1993
• 资助国家: 美国
• 起止时间: 1993-01-01 至 1996-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2222221
• 关键词: antisense nucleic acid; antiserum; biomarker; complementary DNA; epithelium; gene expression; genetic library; genetic regulation; glucose; heat; hypoxia; ion exchange chromatography; mesenchyme; molecular biology; molecular cloning; nucleic acid probes; posttranscriptional RNA processing; posttranslational modifications; protein sequence; protein structure function; pulmonary artery; stress proteins; tissue /cell culture; transfection; ultracentrifugation; vascular endothelium

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.

在体内，生物体及其驻留的组织和细胞经常被 暴露于环境氧气减少的情况。 因此，对急性 缺氧，在某些情况下，适应慢性缺氧，对于 存活率在不同细胞之间存在差异。 一些自适应的 已经提出了允许发展缺氧耐受性的策略 并进行评估；其他人可能不为人知。 我们最近确定了一个 在耐缺氧内皮细胞中诱导的一组特定的应激蛋白 细胞暴露于急性和慢性缺氧期间。 这些蛋白质， 称为缺氧相关蛋白（HAP），是由 环境缺氧，与其他描述的应激蛋白不同， 例如热休克或葡萄糖调节蛋白，并且很可能 在RNA水平上进行调控。 表征内皮细胞 HAP 和 为了更全面地监管，我们将： 1）检查 HAP 的本地化 肺动脉内皮细胞内； 2）调查 HAP 和其他应激蛋白作为保护作用的关系 缺氧、热休克和葡萄糖剥夺的毒性作用； 3）隔离 通过直接蛋白质测序和从缺氧环境中克隆 HAP cDNA 来检测 HAP 刺激肺动脉内皮细胞cDNA文库； 4）使用 分离的蛋白质或从 HAPs cDNA 推导的蛋白质序列 开发分子探针； 5) 使用抗体和 cDNA 探针 研究 HAP 的转录和翻译调控 培养细胞和缺氧组织并确定其效果 改变内皮细胞对缺氧反应的 HAP 表达。 这些 研究将深入了解 HAP 在生物制剂中的调节 和基因水平，并可能提供细胞缺氧的标志物 组织。