Global identification of endogenous ERAD substrates

内源性 ERAD 底物的整体鉴定

• 批准号: 9365628
• 负责人: JAMES A OLZMANN
• 金额: $ 22.53万
• 依托单位: UNIVERSITY OF CALIFORNIA BERKELEY
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-07-01 至 2019-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9365628
• 关键词: ATP phosphohydrolase; Amyloidosis; Biomedical Research; CRISPR/Cas technology; Cell Line; Cell physiology; Cell surface; Cells; Cellular biology; Cholesterol; Communities; Complex; Cytoplasm; Detection; Disease; Dislocations; Endoplasmic Reticulum; Endoplasmic Reticulum Degradation Pathway; Ensure; Enzymes; Fatty Acids; Genes; Goals; Growth Factor; Health; Human; Hydroxymethylglutaryl-CoA reductase; Integral Membrane Protein; Knowledge; Laboratories; Link; Lipids; Liver diseases; Mediating; Membrane; Metabolic; Metabolic Diseases; Methods; Mixed Function Oxygenases; Modeling; Modification; Molecular; Molecular Conformation; Monitor; Mutation; Neurodegenerative Disorders; Pathway interactions; Peptide Hydrolases; Polyubiquitin; Process; Proteins; Proteolysis; Proteome; Proteomics; Quality Control; Reporting; Research; Role; Squalene; Sterols; System; Testing; Ubiquitination; age related; extracellular; fatty acid supplementation; genome editing; improved; inhibitor/antagonist; misfolded protein; overexpression; prevent; protein aggregation; protein degradation; protein folding; proteostasis; receptor; trafficking; ubiquitin-protein ligase

PROJECT SUMMARY / ABSTRACT The long-term goal of our research is to achieve a global and mechanistic understanding of the process by which proteins transiting the endoplasmic reticulum (ER) are recognized and targeted for degradation. The ER is responsible for the folding, modification, and trafficking of approximately one-third of the cellular proteome, including integral membrane proteins that are presented on the cell surface (e.g. channels and receptors) and soluble proteins that are secreted (e.g. growth factors and extracellular proteases). The quality of the proteins that are deployed from the ER is tightly monitored and controlled by a system now known as ER-associated degradation (ERAD). ERAD employs an extensive network of components that mediate misfolded protein detection, dislocation into the cytoplasm, ubiquitination, and proteasomal degradation. In addition to its role in the clearance of misfolded proteins (i.e. quality control), ERAD also regulates the abundance of proteins (i.e. quantity control) to modulate important cell biology processes. For example, ERAD mediates the sterol- dependent degradation of enzymes necessary for cholesterol synthesis, HMG CoA reductase and squalene monooxygenase. Due to the lack of global methods to study ERAD, our understanding of the pools of endogenous substrates that are degraded by different ERAD pathways remains limited. To overcome this obstacle, we developed a VCP-inhibitor substrate trapping approach (VISTA) to identify endogenous ERAD substrates. In our proposed research we will apply this unique strategy to: 1) define pathway-specific ERAD substrates and 2) identify metabolically regulated ERAD substrates. Our studies will characterize a new method of broad utility to the biomedical research community and will advance our understanding of the role of ERAD in cellular protein homeostasis.

项目摘要 /摘要 我们研究的长期目标是通过通过 转移内质网（ER）的蛋白质被识别并靶向降解。急诊室 负责大约三分之一的细胞蛋白质组折叠，修饰和运输， 包括在细胞表面（例如通道和受体）和 分泌的可溶性蛋白质（例如生长因子和细胞外蛋白酶）。蛋白质的质量 从急诊室中部署的系统由现在称为ER相关的系统严格监控和控制 退化（Erad）。 Erad采用了广泛的组件网络，可以介导错误折叠的蛋白质 检测，脱位到细胞质，泛素化和蛋白酶体降解。除了它在 错误折叠蛋白的清除（即质量控制）也调节了蛋白质的丰度（即 数量控制）调节重要的细胞生物学过程。例如，Erad介导了固醇 胆固醇合成，HMG COA还原酶和小甲烯的依赖性降解酶的降解 单加氧酶。由于缺乏研究ERAD的全球方法，我们对池的理解 由不同的ERAD途径降解的内源性底物仍然有限。克服这一点 障碍物，我们开发了VCP抑制剂底物陷阱方法（VISTA）以识别内源性ERAD 基材。在我们拟议的研究中，我们将把这种独特的策略应用于：1）定义特定途径的ERAD 底物和2）确定代谢调节的ERAD底物。我们的研究将描述一个新的 对生物医学研究社区的广泛实用性方法，并将促进我们对 细胞蛋白稳态中的ERAD。