Engagement and Communication Between Proteasonal Subcomplexes

蛋白酶子复合物之间的参与和交流

• 批准号: 9892109
• 负责人: ROBERT JOSEPH TOMKO
• 金额: $ 9.38万
• 依托单位: FLORIDA STATE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-04-01 至 2022-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9892109
• 关键词: ATP Hydrolysis; ATP phosphohydrolase; Active Sites; Affect; Alzheimer' s Disease; Antineoplastic Agents; Architecture; Area; Binding; Biochemical Genetics; Biogenesis; Biological Assay; Biology; Bortezomib; Catalysis; Cell Cycle; Cells; Cellular biology; Chemicals; Cleaved cell; Clinic; Communication; Complex; Coupled; Coupling; DNA Repair; Deubiquitinating Enzyme; Diabetes Mellitus; Disease; Drug resistance; Elements; Event; Garbage; Goals; Heart; Human; In Vitro; Individual; Inflammatory; Link; Macromolecular Complexes; Malignant Neoplasms; Mediating; Molecular; Molecular Biology; Molecular Conformation; Movement; Mutation; Neurodegenerative Disorders; Non-Insulin-Dependent Diabetes Mellitus; Nucleosome Core Particle; Nucleotides; Patients; Peptides; Process; Proteasome Inhibition; Proteasome Inhibitor; Proteins; Proteolysis; Recombinants; Reporter; Research; Role; Route; Signal Transduction; Structure; Testing; Velcade; Work; adaptive immunity; base; combinatorial; drug development; genetic approach; high throughput screening; human disease; in vivo; insight; multicatalytic endopeptidase complex; novel; novel therapeutics; protein degradation; protein misfolding; small molecule; small molecule inhibitor; tool

Project Summary/Abstract The proteasome is a multisubunit macromolecular machine that mediates most regulatory protein degradation and removes toxic proteins from cells. It is essential for activities as diverse as the cell cycle, adaptive immunity, and DNA repair. Alterations to proteasome activity impact numerous human diseases, including cancer, neurodegenerative disorders, and diabetes. The proteasome consists of three functional subcomplexes: the lid, the base, and the core particle. Each subcomplex performs distinct functions during substrate degradation. The lid removes the proteasomal targeting signal, the base uses energy from ATP hydrolysis to unfold the substrate, and the core particle then cleaves it into short peptides. These activities—as well as the subcomplexes that harbor them—are intimately linked by static and dynamic inter-subcomplex interactions. Recent structural studies have unexpectedly revealed that the proteasome exists in at least two well-defined conformational states—an apo state, in which the substrate passageways and the enzymatic active sites within these subcomplexes are blocked, and an engaged state, in which these passageways and active sites are opened and aligned, ready to accept and process substrates. Thus, these states reflect “off” and “on” conformations for the proteasome, respectively. Proteolytic inhibitors of the proteasome such as Velcade (bortezomib) are proven anticancer drugs, but resistance to these agents is already emerging. This necessitates alternative approaches to control proteasome function. Manipulation of the conformational state of the proteasome could allow for their selective activation or inactivation at will. This strategy could permit treatment of proteasome-addicted cancers via proteasome inactivation, as well as treatment of proteinopathies such as Alzheimer’s and type II diabetes, via enhancement of proteolysis to clear toxic inclusions. The long-term goal of this project is to understand the molecular mechanisms regulating engagement and communication between proteasomal subcomplexes, and how they relate to the proteasome’s conformational state. We seek to determine how individual conformation- specific contacts between lid and base subunits control proteasome structure and function (Aim1), dissect the critical role of nucleotide binding in reorganization of the lid-base interface to promote the engaged state (Aim 2), and to develop small molecules that disrupt lid-base coordination to be used as tools for studying proteasome function in human cells or in vitro (Aim 3). We anticipate our studies will yield insights into allosteric communication, energy use, and substrate processing by the proteasome, as well as yielding new information on proteasome biogenesis and structure. Further, the functional elements of the proteasome are found in many other multiprotein machines, so our studies could reveal general principles governing the function of diverse macromolecular complexes, and thus will impact numerous areas of cell and molecular biology.

项目摘要/摘要 蛋白酶体是一种多生育大分子机器，可介导大多数调节蛋白 降解并去除细胞中的有毒蛋白质。对于像细胞周期一样潜水的活动至关重要， 自适应免疫学和DNA修复。蛋白酶体活动的改变会影响许多人类疾病， 包括癌症，神经退行性疾病和糖尿病。蛋白酶体由三个功能组成 子复合：盖子，底座和核心粒子。每个子复合在期间执行不同的功能 底物降解。盖子去除蛋白酶体靶向信号，基数使用ATP的能量 水解以展开底物，然后将其切割成短肽。这些活动 - 如 以及藏有它们的子复合物 - 与静态和动态互化的相关联系在一起 互动。最近的结构研究意外揭示了蛋白酶体至少存在于两个 定义明确的构象状态 - 一个APO状态，底物语音和酶活性 这些子复合物中的站点被阻塞，并且是一个参与的状态，其中这些通道和活跃 打开和对齐站点，准备接受和处理基板。这些国家反映了“关闭”和“ on” 蛋白酶体的构象。 蛋白酶体的蛋白水解抑制剂，例如Velcade（Bortezomib）已被证明是抗癌药物，但 对这些药物的抵抗已经出现。这种控制蛋​​白酶体的必要替代方法 功能。操纵蛋白酶体的构象状态可以允许其选择性激活或 随意失活。该策略可以允许通过蛋白酶体治疗蛋白酶体成瘾的癌症 通过增强，灭活以及诸如阿尔茨海默氏症和II型糖尿病等蛋白质病的治疗 蛋白水解以清除有毒的夹杂物。该项目的长期目标是了解分子 应对蛋白酶体子复合物之间的参与和沟通的机制，以及如何 它们与蛋白酶体的构象状态有关。我们试图确定个人构象如何 盖子和基部亚基之间的特定接触控制蛋白酶体的结构和功能（AIM1），剖析 核苷酸结合在盖碱界面重组以促进参与状态的关键作用（AIM 2），并开发出破坏盖碱配位的小分子，以用作研究蛋白酶体的工具 在人类细胞或体外功能（AIM 3）。我们预计我们的研究将产生对变构的见解 蛋白酶体的沟通，能源使用和底物处理，并产生新信息 关于蛋白酶体的生物发生和结构。此外，在许多人中发现了蛋白酶体的功能元素 其他多蛋白机器，因此我们的研究可以揭示有关潜水员功能的一般原则 大分子复合物，因此会影响细胞和分子生物学的许多领域。