Molecular Mechanism and In Vivo Study of ER Associated Degradation

ER 相关降解的分子机制和体内研究

• 批准号: 8321725
• 负责人: Lenka Kundrat
• 金额: $ 5.39万
• 依托单位: WHITEHEAD INSTITUTE FOR BIOMEDICAL RES
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-08-01 至 2013-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8321725
• 关键词: B-Lymphocytes; Cells; Crosslinker; Cytosol; Development; Disease; Dislocations; Dominant-Negative Mutation; Enzymes; Excision; Future; Generations; Goals; Knowledge; Liver; Mediating; Membrane; Methodology; Molecular; Organism; Positioning Attribute; Principal Investigator; Process; Proteins; Quality Control; Reaction; Stress; Tail; Techniques; Tissues; Ubiquitin; Ubiquitin-Conjugating Enzymes; Ubiquitination; base; career development; in vivo; mouse model; mutant; novel; protein complex; protein misfolding; research study; response; small hairpin RNA; sortase; tool; transpeptidation; ubiquitin ligase; ubiquitin-protein ligase; ubiquitin-specific protease

DESCRIPTION (provided by applicant): Protein quality control entails the removal of misfolded proteins. Accumulation of such misfits leads to activation of the unfolded protein response (UPR) with eventual pathological consequences to the cell and organism, should misfolded proteins persist. Proteins, either luminal or membrane-disposed, that fail to fold in the ER are often retro-translocated or 'dislocated' into the cytosol for degradation. This dislocation is to a large extent ubiquitin- dependent, as is the degradation of misfolded cytosolic proteins. The overall goal of the project is to study the mechanism of ER associated degradation (ERAD) and how disruption of this process leads to, or is an important factor in, the development of various protein misfolding diseases. To achieve this goal, novel catch-and-release probes will be applied in combination with photo-crosslinkers to identify new components involved in dislocation: in vivo ubiquitin acceptors of the ERAD ubiquitinating conjugating enzyme Ubc6e and in vivo interaction partners of Ubc6e. It is currently difficult to extend the study of ER stress control to the organismal level. The Ploegh lab has developed tools that now allow us to block the ER dislocation process in a non-toxic manner, an approach that will be exploited and extended to two new in vivo mouse models. Identification of novel dislocation components will be an important extension of our basic knowledge of ERAD machinery. Because many diseases have been attributed to the accumulation of misfolded proteins, the proposed experiments are relevant to numerous diseases.

描述（由申请人提供）：蛋白质质量控制需要去除错误折叠的蛋白质。这种错配的积累导致未折叠蛋白反应（UPR）的激活，如果错误折叠的蛋白持续存在，最终会对细胞和生物体造成病理后果。在内质网中不能折叠的蛋白质，无论是腔内还是膜内处理的，通常会被反向易位或“错位”到细胞质中进行降解。这种错位在很大程度上依赖于泛素，就像错误折叠的细胞质蛋白的降解一样。该项目的总体目标是研究内质网相关降解（ERAD）的机制，以及该过程的破坏如何导致各种蛋白质错误折叠疾病的发生，或者是其重要因素。为了实现这一目标，新的捕获-释放探针将与光交联剂结合使用，以识别参与错位的新组分：ERAD泛素化偶联酶Ubc6e的体内泛素受体和Ubc6e的体内相互作用伙伴。目前很难将内质网应激控制的研究扩展到机体水平。Ploegh实验室已经开发出工具，现在允许我们以无毒的方式阻断内质网错位过程，这种方法将被利用并扩展到两种新的体内小鼠模型中。识别新的位错成分将是我们对ERAD机械基础知识的重要扩展。由于许多疾病归因于错误折叠蛋白质的积累，因此提出的实验与许多疾病相关。