Visualizing the Mechanisms of Protein Quality Control

蛋白质质量控​​制机制的可视化

• 批准号: 10164810
• 负责人: Peter Shen
• 金额: $ 38.13万
• 依托单位: UNIVERSITY OF UTAH
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-08-01 至 2024-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10164810
• 关键词: ATP Hydrolysis; ATP phosphohydrolase; Address; Autophagocytosis; Binding; Cell Cycle Progression; Cell physiology; Cells; Clinical; Complex; Cryoelectron Microscopy; Degenerative Disorder; Enzymes; Equilibrium; Functional disorder; Gene Expression; Goals; Human; Image; Inclusion Body Myopathy with Early-Onset Paget Disease; Life; Malignant Neoplasms; Membrane Fusion; Methods; Molecular; Molecular Conformation; Molecular Machines; Mutation; Pathway interactions; Protein Biosynthesis; Proteins; Proteomics; Quality Control; Research; Role; Structure; Therapeutic; Work; cancer therapy; cofactor; familial amyotrophic lateral sclerosis; inhibitor/antagonist; insight; programs; tool; trafficking; tumor; valosin-containing protein

Project Summary / Abstract Optimal cellular function requires balanced networks that maintain protein synthesis, folding, trafficking, remodeling and degradation. The Cdc48/p97/VCP AAA ATPase is an essential and abundant molecular machine that helps maintain this balance across eukaryotic life and is a critical control point for many cellular pathways. Cdc48 is best characterized for its role in targeting ubiquitylated proteins for proteasomal degradation, but the enzyme also functions in a wide range of other essential pathways, including cell cycle progression, autophagy, membrane fusion, and gene expression. Mutations in human Cdc48 cause a multisystem proteopathy that clinically manifests as a combination of Inclusion Body Myopathy, Paget's Disease of Bone, Frontotemporal Dementia (collectively known as IBMPFD), and familial Amyotrophic Lateral Sclerosis (fALS). Moreover, Cdc48 expression is elevated in several tumors and its inhibitors are an emerging class of therapeutics for cancer treatment. Despite these critical roles, surprisingly little is known about the molecular mechanisms that allow Cdc48 to perform its myriad cellular functions. More than thirty adaptors and cofactors are known to interact with Cdc48, but how cells organize these binding partners into functional complexes is not well understood. My research program aims to define the mechanisms underlying Cdc48 functions by using an integrative approach of endogenous purification, proteomics, cryo-EM imaging, and computational processing methods to visualize and characterize Cdc48 assemblies in an array of its native compositional and conformational states. During the project period, we propose to achieve two major goals: first, we aim to determine the structures of native Cdc48 assemblies and their molecular determinants that functionally separate the enzyme across multiple cellular pathways; second, we seek to resolve how Cdc48 converts energy from ATP hydrolysis into a pulling force that remodels and unfolds its protein substrates. Addressing these questions is essential to understand how Cdc48 drives a wide range of cellular processes and provide insights into how its dysfunction and misregulation contribute to degenerative disease and cancer. The tools we develop to accomplish these goals will likely be broadly applicable in defining the structural landscapes of other challenging molecular machines.

项目摘要 /摘要 最佳细胞功能需要平衡网络，以维持蛋白质合成，折叠，运输， 重塑和降解。 Cdc48/p97/vcp AAA ATPase是必不可少的分子 有助于维持真核生活平衡的机器，这是许多蜂窝的关键控制点 途径。 CDC48最能以其在靶向泛素化蛋白中的蛋白酶体方面的作用。 降解，但该酶在其他各种必需途径中也起作用，包括细胞周期 进展，自噬，膜融合和基因表达。人类CDC48中的突变导致 临床上表现为包容性身体肌病的组合，Paget的多系统蛋白质病 骨骼疾病，额颞痴呆（统称为IBMPFD）和家族性肌萎缩性侧面 硬化症（伪造）。此外，在几种肿瘤中CDC48表达升高，其抑制剂是出现的 用于癌症治疗的治疗剂类。尽管这些关键作用，但令人惊讶的是 允许CDC48执行其无数细胞功能的分子机制。超过三十个适配器， 已知辅因子与CDC48相互作用，但是细胞如何将这些结合伙伴组织到功能上 复合体并不理解。我的研究计划旨在定义CDC48的基础机制 通过使用内源性纯化，蛋白质组学，冷冻EM成像和 计算处理方法可视化和表征其本机数组中的Cdc48组件 组成和构象状态。在项目期间，我们建议实现两个主要目标： 首先，我们旨在确定天然CDC48组件及其分子决定因素的结构，即 在功能上将酶跨多个细胞途径分开；其次，我们寻求解决CDC48的方式 将ATP水解的能量转化为重塑并展开其蛋白质底物的拉力。 解决这些问题对于了解CDC48如何驱动各种蜂窝过程至关重要 并提供有关其功能障碍和错误调节如何导致退化性疾病和癌症的见解。 我们开发的来实现这些目标的工具可能广泛地适用于定义结构 其他具有挑战性的分子机器的景观。