ROLES OF OST3/6 THIOL OXIDOREDUCTASES IN N-LINKED PROTEIN GLYCOS/REDOX HOMEOSTAS

OST3/6 硫醇氧化还原酶在 N-连接蛋白糖/氧化还原稳态中的作用

• 批准号: 8168314
• 负责人: Dmitri Fomenko
• 金额: $ 14.17万
• 依托单位: UNIVERSITY OF NEBRASKA LINCOLN
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-08-01 至 2011-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8168314
• 关键词: Address; Biological Assay; Biological Process; Brain Diseases; Colorectal; Complex; Computer Retrieval of Information on Scientific Projects Database; Enzymes; Eukaryota; Family; Funding; Gene Expression; Goals; Grant; Homeostasis; Human; Institution; Knock-out; Knockout Mice; Link; Malignant Neoplasms; Malignant neoplasm of pancreas; Mammalian Cell; Mediating; Membrane; Mental Retardation; Metastatic Prostate Cancer; Methods; Modeling; Modification; N33 gene; Oxidation-Reduction; Oxidative Stress; Oxidoreductase; Process; Protein Deficiency; Protein Glycosylation; Proteins; Regulation; Research; Research Personnel; Resources; Role; Saccharomyces cerevisiae; Series; Signal Transduction; Source; Stress; Sulfhydryl Compounds; Thioredoxin; Tumor Suppressor Proteins; United States National Institutes of Health; Yeasts; disulfide bond; dolichyl-diphosphooligosaccharide - protein glycotransferase; glycosylation; link protein; mouse model; protein folding; research study; tool

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. The overall goal of this project is to provide understanding of the roles of OST3/6 thiol oxidoreductases in Nlinked protein glycosylation and ER redox homeostasis. Thiol-dependent redox processes are involved in oxidative stress defense, signal transduction, and protein folding, modification and regulation, and are catalyzed by structurally distinct families of enzymes known as thiol oxidoreductases. Numerous ER thiol oxidoreductases involved in protein folding have been characterized; however, the overall machinery of folding and glycosylation remains poorly characterized. The N-linked protein glycosylation in the ER is an essential process and a key step in the control of protein folding in eukaryotes. OST3/6 proteins are abundant ER membrane-linked thioredoxin-fold thiol oxidoreductases involved in the redox control of N-linked protein glycosylation in the oligosaccharyltransferase complex. OST3/6 deficiency is associated with severe protein underglycosylation and ER stress. Homozygous deletion of human OST3/6 like protein, N33, correlates with metastatic prostate cancer and its allelic deletion is associated with human colorectal and pancreatic cancers. This observation suggests a possible tumor suppressor function of N33. In addition, there are two known cases of a natural knockout of N33 in humans which are associated with nonsyndromic mental retardation. In the proposed study, we will systematically characterize the biological function of OST3/6 proteins. The effect of OST3/6 deficiency will be examined with regard to efficiency of protein glycosylation and ER stress. Possible targets of OST3/6 proteins will be identified using thiol-mediated substrate-trapping method and global gene expression analysis. The OST3/6 roles in disulfide bond formation will be addressed in series of thiol oxidoreductase assays. These experiments will be carried in yeast Saccharomyces cerevisiae and mammalian cells. We also would like to develop OST3/6 knockout mouse models. These models will provide tools in a better understanding of the consequences of OST3/6 protein deficiency on cancer and brain disorders and will be useful in the analysis of OST3/6 biological function.

该副本是利用众多研究子项目之一 由NIH/NCRR资助的中心赠款提供的资源。子弹和 调查员（PI）可能已经从其他NIH来源获得了主要资金， 因此可以在其他清晰的条目中代表。列出的机构是 对于中心，这不一定是调查员的机构。 该项目的总体目标是提供对NLinked中OST3/6硫醇氧化酶的作用的理解 蛋白质糖基化和ER氧化还原稳态。硫醇依赖性氧化还原过程参与 氧化应激防御，信号转导和蛋白质折叠，修饰和调节，并且是 被称为硫醇氧化酶的酶的结构不同的酶催化。许多ER硫醇 已经表征了参与蛋白质折叠的氧化还原酶。但是，折叠的整体机制 糖基化的特征仍然很差。 ER中的N连锁蛋白糖基化是必不可少的 控制真核生物中蛋白质折叠的过程和关键步骤。 OST3/6蛋白丰富ER 膜连接的硫氧还蛋白折叠的硫氧还氧化还原酶参与N-连接蛋白的氧化还原对照 寡糖胆碱转移酶复合酶中的糖基化。 OST3/6缺陷与严重蛋白质有关 糖基化和ER应力太多。人ost3/6的纯合缺失，例如蛋白质，N33，与 前列腺癌及其等位基因缺失与人类结肠直肠和胰腺癌有关。 该观察结果表明N33可能具有肿瘤抑制功能。此外，有两种已知情况 人类中N33的自然敲除，与非智力智力低下有关。在 拟议的研究，我们将系统地表征OST3/6蛋白的生物学功能。效果 关于蛋白质糖基化和ER应激的效率，将检查OST3/6缺乏症。可能的 将使用硫醇介导的底物捕捞方法和全局基因鉴定OST3/6蛋白的靶标 表达分析。二硫键形成中的OST3/6角色将在一系列硫醇中解决 氧化还原酶测定。这些实验将在酿酒酵母和哺乳动物的酵母菌中进行 细胞。我们还想开发OST3/6基因敲除鼠标模型。这些模型将在 更好地理解OST3/6蛋白缺乏对癌症和脑部疾病的后果，并将 在OST3/6生物学功能的分析中有用。