Engagement and Communication Between Proteasomal Subcomplexes

蛋白酶体亚复合物之间的参与和通讯

• 批准号: 10659372
• 负责人: ROBERT JOSEPH TOMKO
• 金额: $ 30.16万
• 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 2027-02-28
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10659372
• 关键词: 26S proteasome; ATP phosphohydrolase; Animals; Architecture; Area; Autoimmune Diseases; Autophagocytosis; Binding; Biogenesis; Biological Assay; Biology; Budgets; Catalysis; Cells; Communicable Diseases; Communication; Complement; Complex; Conserved Sequence; Coupled; Cytoprotection; Dedications; Diabetes Mellitus; Disease; Dissociation; Distant; Drug Targeting; Elements; Equilibrium; Eukaryota; Event; Excision; Family; Fluorescence Resonance Energy Transfer; Goals; Health; Hot Spot; Human; Infection; Kinetics; Knowledge; Ligand Binding; Ligands; Link; Malignant Neoplasms; Mediating; Microsporidia; Modeling; Molecular; Molecular Chaperones; Molecular Conformation; Movement; Mutation; Nerve Degeneration; Neurodegenerative Disorders; Nucleosome Core Particle; Parasites; Parasitic infection; Pharmaceutical Preparations; Process; Proteins; Proteolysis; Quality Control; Reagent; Recycling; Role; Route; Signal Transduction; Site; Structure; Testing; Tyrosine; United States National Institutes of Health; Work; Yeasts; anti-cancer; base; combat; conformational conversion; design; drug discovery; effective therapy; flexibility; guided inquiry; human disease; human pathogen; in vivo; inhibitor; insight; interest; multicatalytic endopeptidase complex; novel; novel therapeutics; priority pathogen; protein degradation; tool; transmission process

Project Summary Abstract The 26S proteasome conducts most regulated protein degradation and eliminates toxic proteins from cells. The proteasome is a validated anti-cancer target, and holds substantial promise as a target for treatment of neurodegenerative disorders and some infectious diseases. Our long-term goal is to understand how the three major complexes of the proteasome—the lid, base, and core particle—engage and communicate within and between one another. We have thus far made significant progress toward this goal and have developed a number of novel tools and reagents that have furthered our understanding of intra- and inter-complex communication. Conceptual advances have included: i) discovery of additional conformational states of the yeast proteasome relevant to substrate catalysis; ii) demonstration that six highly similar ATP-hydrolyzing subunits differentially influence the activation state of the proteasome; iii) discovery of a link between the conformational state of the proteasome and release of a dedicated proteasome assembly chaperone; and iv) and the finding that proteasomal subcomplexes disengage one another prior to their destruction by autophagy. A paradigm emerging from this initial budget period is that rather small binding events or molecular movements are transmitted, often over long distances, to enact largescale conformational changes. Understanding how such local events are amplified and transmitted to distant areas of the proteasome to coordinate assembly and catalysis is thus a critical knowledge gap. In this first renewal, we propose three Aims that explore examples of this paradigm newly discovered by us during the initial budget period. Together, they will push our knowledge of proteasome dynamics and inter-complex communication into new arenas. In the first, we will use newly developed FRET-based kinetic assays to decipher how local changes to the lid-base interface regulate the timely binding and release of dedicated assembly chaperones from nascent proteasomes. In the second, we will explore a surprising allosteric conduit originating from the substrate unfolding center of the proteasome that regulates the stability between two key subcomplexes. In the third Aim, we will investigate an unusual eukaryotic proteasome from a poorly studied human parasite from the phylum of Microsporidia. Microsporidia lack several proteasome subunits that normally span a key inter-complex interface. The missing subunits contain several small sequence elements with essential roles in assembly and catalysis in other eukaryotes, so exploring these unusual proteasomes will thus reveal both conserved and unique elements of inter-complex communication. These studies are anticipated to produce important insights into the engagement and communication between the proteasomal subcomplexes, significantly advancing several aspects of proteasome biology and drug discovery. Further, microsporidia are NIH priority pathogens of interest for which no broadly effective treatments exist. Insights into microsporidial proteasomes thus may reveal new targets to combat certain parasitic infections.

项目摘要摘要 26S蛋白质组会进行大多数调节的蛋白质降解，并消除细胞中的有毒蛋白质。这 蛋白酶体是一个经过验证的抗癌靶 神经退行性疾病和一些传染病。我们的长期目标是了解三个 蛋白酶体的主要复合物（盖，底部和核心粒子）在内部和沟通 彼此之间。到目前为止，我们已经朝着这个目标取得了重大进展，并发展了一个数字 新型工具和试剂的新工具和试剂进一步了解了复杂性交流。 概念的进步包括：i）发现酵母蛋白酶体的其他构象状态 与底物催化剂有关； ii）证明了六个高度相似的ATP - 水解亚基差异 影响蛋白酶体的激活状态； iii）发现构象状态之间的联系 蛋白酶体和专用蛋白酶体组装伴侣的释放；和iv）以及发现 蛋白酶体子复合物在自噬毁灭之前相互脱离。 从最初的预算期开始出现的范式是相当小的结合事件或分子运动 通常在长距离上传输，以实现大规模构象变化。了解如何这样 将当地事件放大并传输到蛋白酶体的遥远区域，以协调组装和 因此，催化是一个关键的知识差距。在第一次续约中，我们提出了三个探索示例的目标 我们在最初的预算期间新发现的这个范式。他们在一起将推动我们对 蛋白酶体的动力学和复合沟通与新领域的交流。首先，我们将使用新的 开发了基于FRET的动力学测定，以解读局部变化对盖碱接口的变化如何调节及时的 来自新生蛋白酶体的专用组装伴侣的结合和释放。在第二个中，我们将 探索源自蛋白酶体的底物展开中心的令人惊讶的变构管道 调节两个关键子复合物之间的稳定性。在第三个目标中，我们将研究一个异常的真核性 来自微孢子虫门的人类寄生虫的蛋白酶体。微孢子虫缺乏几种 通常跨越关键复合体接口的蛋白酶体亚基。缺失的亚基包含几个 小序列元素在其他真核生物中具有基本作用和催化作用，因此探索这些 因此，不寻常的蛋白酶体将揭示复杂交流的保守和独特元素。 预计这些研究将对参与和交流之间的重要见解 蛋白酶体子复合物，显着推进了蛋白酶体生物学和药物的几个方面 发现。此外，微孢子虫是NIH优先病原体，没有广泛有效的治疗方法 存在。因此，对微孢子虫蛋白酶体的洞察力可能会揭示出应对某些寄生虫感染的新靶标。