“Regulation of Protein Quality Control by the VCP AAA-ATPase”

– VCP AAA-ATPase 对蛋白质质量控制的调节 –

基本信息

批准号：
10598249
负责人：
Malavika Raman
金额：
$ 1.86万
依托单位：
TUFTS UNIVERSITY BOSTON
依托单位国家：
美国
项目类别：
财政年份：
2018
资助国家：
美国
起止时间：
2018-05-01 至 2024-03-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10598249
关键词：
ATP phosphohydrolase Adaptor Signaling Protein Affect Aging Alzheimer&apos s Disease Amyotrophic Lateral Sclerosis Animal Model Autophagocytosis Binding Cell Aging Cells Cellular Stress Cellular Structures Cessation of life Clinical Complex Degradation Pathway Development Disease Doctor of Philosophy Funding Future Goals Homeostasis Huntington Disease Inclusion Body Myopathy with Early-Onset Paget Disease Individual K22 Award Knock-out Link Mediating Molecular Chaperones Molecular Target Mutate Mutation Nerve Degeneration Neurodegenerative Disorders Neurons Normal Cell Organelles Outcome Parkinson Disease Pathway interactions Play Prion Diseases Process Protein translocation Proteins Proteome Proteomics Quality Control Regulation Role Site Stress Structure System Testing Therapeutic Toxic effect Translating Ubiquitin base biological adaptation to stress cellular targeting cytotoxic endoplasmic reticulum stress glycosylation high resolution imaging member misfolded protein multicatalytic endopeptidase complex mutant neuron loss new therapeutic target prevent protein aggregation protein complex protein folding protein function proteostasis recruit response small molecule valosin containing protein mutation valosin-containing protein

项目摘要

Project Abstract Protein quality control is of critical importance in maintaining cellular protein homeostasis, especially during stress, disease states and aging. These networks are comprised of: 1) chaperones that assist in protein folding and stabilization of intermediates to prevent aggregation, and 2) degradation pathways, such as the ubiquitin proteasome system and autophagy, to eliminate proteins and organelles with sub-optimal function. A decline in protein quality control in aging cells contributes to the development of numerous genetically and clinically distinct neurodegenerative disorders such as amyotrophic lateral sclerosis (ALS), Alzheimer's, Parkinson's, Huntington's and prion diseases. Valosin containing protein (VCP, also known as p97) is a AAA-ATPase that plays an important role in the ubiquitin proteasome system to capture ubiquitylated substrates via designated adaptors to mediate substrate degradation. Mutation of VCP causes several neurodegenerative disorders including ALS and a rare multi-system disorder, inclusion body myopathy, Paget’s disease of the bone and frontotemporal dementia (IBMPFD). The most commonly mutated region of VCP resides at the site of adaptor binding and in certain instances these mutations alter the constellation of adaptors that are bound to VCP skewing its targeting to substrates. A common cellular feature of VCP mutations is the formation and persistence of protein aggregates that fail to be cleared. We have identified a specific VCP-adaptor complex that is targeted to the aggresome, a structure that sequesters misfolded aggregated proteins formed during cell stress. While aggresomes are generally cyto-protective, under conditions where they persist, for example in aging neurons with declining protein quality control pathways, they eventually become cytotoxic. Depletion of the VCP adaptor leads to a deficit in aggresome formation, stabilization of ubiquitylated substrates and triggers ER stress. We hypothesize that VCP mediates aggresome clearance by adaptor specific targeting to enable clearance of specific misfolded or aggregated substrates. We will use a combination of targeted molecular studies, quantitative proteomics and high-resolution imaging to achieve the main objectives of this proposal. We will (1) characterize the VCP aggresome targeted complex to determine mechanisms of targeting and its functions within the aggresome (2) identify ubiquitylated cellular targets of the VCP complex targeted to the aggresome using ubiquitin remnant capture proteomics and (3) determine whether disease-relevant VCP mutations impact its association with the aggresome complex and test if the VCP adaptor can recognize and enable clearance of disease relevant protein aggregates. Successful implementation of this proposal will identify new targets that can be pursued for the development of agents that enable aggregate clearance in a number of neurodegenerative disorders.

项目摘要蛋白质质量控制对于维持细胞蛋白质稳态至关重要，尤其是在这些网络由以下部分组成：1) 协助蛋白质折叠的分子伴侣。和稳定中间体以防止聚集，以及 2) 降解途径，例如泛素蛋白酶体系统和自噬，消除功能次优的蛋白质和细胞器。衰老过程中的蛋白质质量控制有助于细胞发育许多遗传和临床上独特的细胞神经退行性疾病，如肌萎缩侧索硬化症 (ALS)、阿尔茨海默病、帕金森病、亨廷顿舞蹈症含 Valosin 的蛋白（VCP，也称为 p97）是一种 AAA-ATP 酶，可发挥作用。在泛素蛋白酶体系统中通过指定的接头捕获泛素化底物的重要作用 VCP 突变会介导底物降解，导致多种神经退行性疾病，包括 ALS 和一种罕见的多系统疾病、包涵体肌病、佩吉特骨病和额颞叶痴呆 (IBMPFD) VCP 最常见的突变区域位于接头结合位点和某些部位。在这种情况下，这些突变改变了与 VCP 结合的适配器群，使其靶向偏向于 VCP 突变的一个常见细胞特征是蛋白质聚集体的形成和持续存在。我们已经确定了一种针对攻击性物质的特定 VCP 适配器复合物。隔离在细胞应激过程中形成的错误折叠聚集蛋白的结构。在它们持续存在的条件下，例如在衰退的衰老神经元中，通常具有细胞保护作用蛋白质质量控制途径，它们最终会产生细胞毒性。VCP 接头的耗尽会导致细胞毒性。攻击体形成、泛素化底物稳定的缺陷并引发内质网应激。 VCP 通过适配器特异性靶向介导攻击性清除，从而能够清除特定的错误折叠我们将结合使用靶向分子研究、定量蛋白质组学和高分辨率成像以实现本提案的主要目标，我们将 (1) 描述 VCP 的特征。攻击体靶向复合体，用于确定攻击体中的靶向机制及其功能 (2) 使用泛素残留物识别针对攻击体的 VCP 复合物的泛素化细胞靶标捕获蛋白质组学并 (3) 确定与疾病相关的 VCP 突变是否影响其与疾病的关联聚集体复合物并测试 VCP 接头是否可以识别并清除疾病相关蛋白该提案的成功实施将确定可实现的新目标。开发能够清除多种神经退行性疾病的药物。