“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：
9884781
负责人：
Malavika Raman
金额：
$ 34.65万
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 2023-03-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9884781
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 Estrogen receptor positive 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稳态至关重要，尤其是在压力，疾病状态和衰老。这些网络的完成：1）有助于蛋白质折叠的伴侣和中间体的稳定以防止聚集，以及2）降解途径，例如泛素蛋白酶体系统和自噬，以消除具有亚最佳功能的蛋白质和细胞器。下降衰老细胞中的蛋白质质量控制有助于发展许多遗传和临床上不同的神经退行性疾病，例如肌萎缩性侧面硬化症（ALS），阿尔茨海默氏症，帕金森氏症，亨廷顿的疾病和病毒疾病。含有蛋白质的瓣膜（VCP，也称为p97）是一种AAA-ATPase 在泛素蛋白酶体系统中的重要作用，以通过指定的适配器捕获泛素化底物介导底物降解。 VCP的突变引起几种神经退行性疾病，包括ALS和一种罕见的多系统疾病，纳入身体肌病，骨骼和额颞痴呆症的paget病（IBMPFD）。在适配器结合部位的VCP住宅最常见的区域，在某些地方实例这些突变改变了与VCP束缚其靶向的适配器的星座基材。 VCP突变的常见细胞特征是蛋白质聚集体的形成和持久性那无法清除。我们已经确定了针对agresome的特定VCP-Audaptor复合物结构隔离在细胞应激期间形成的杂种聚集的蛋白质。虽然是混合物通常，在持续存在的条件下，细胞保护的影响，例如在衰老的神经元下降蛋白质质量控制途径，有时会变成细胞毒性。 VCP适配器的耗竭导致阿格雷斯形成的赤字，泛素化底物的稳定和触发ER应力。我们假设 VCP通过适配器特定的靶向介导了Agresome的清除，以使特定错误折叠或聚合的底物。我们将结合靶向分子研究，定量蛋白质组学和高分辨率成像以实现该提案的主要目标。我们将（1）表征VCP Agresome靶向复合物，以确定靶向机理及其在Agresome中的功能（2）使用泛素残余捕获蛋白质组学，（3）确定与疾病相关的VCP突变是否影响其与 Agresome复合物并测试VCP适配器是否可以识别并能够清除疾病相关蛋白聚合。该提案的成功实施将确定可以追求的新目标开发能够在许多神经退行性疾病中汇总清除的药物。