Understanding Mycobacterium tuberculosis 20S proteasome assembly

了解结核分枝杆菌 20S 蛋白酶体组装

• 批准号: 10689119
• 负责人: Leonila Lagunes
• 金额: $ 7.18万
• 依托单位: UNIVERSITY OF CALIFORNIA LOS ANGELES
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-09-30 至 2024-09-29
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10689119
• 关键词: Actinobacteria class; Affect; Anti-Bacterial Agents; Architecture; Bacteria; Bacterial Infections; Binding; Biochemical; Biological Assay; Biological Phenomena; Biophysics; Cell Cycle Regulation; Cell Survival; Cells; Cellular biology; Complex; Computational Biology; Computer software; Cryoelectron Microscopy; Development; Diffusion; Dimerization; Disease; Drug Targeting; Environment; Eukaryota; Fellowship; Foundations; Future; Goals; Growth; Hour; Human; Immune system; In Vitro; Individual; Innovative Therapy; Kinetics; Knowledge; Life; Macrophage; Mass Spectrum Analysis; Measures; Mentors; Methods; Mission; Modeling; Molecular Structure; Mycobacterium tuberculosis; Nucleosome Core Particle; Organism; Pathway interactions; Patients; Play; Process; Protein Conformation; Proteins; Public Health; Research; Research Personnel; Resistance; Resolution; Rhodococcus; Role; Structure; System; Technical Expertise; Techniques; Testing; Thermodynamics; Time; Training; Tuberculosis; United States; United States National Institutes of Health; Work; biophysical model; career; chemical reaction; clinical application; dimer; drug development; experience; experimental analysis; experimental study; genetic regulatory protein; human disease; immune resistance; improved; inhibitor; innovation; insight; interdisciplinary approach; latent infection; macromolecule; mathematical analysis; mathematical model; misfolded protein; monomer; multicatalytic endopeptidase complex; novel; particle; physical property; predictive modeling; protein degradation; reaction rate; skills; theories; tuberculosis treatment

PROJECT SUMMARY Tuberculosis affects over 8,000 individuals in the United States every year, with Mycobacterium tuberculosis (Mtb) able to resist the immune system in part through proteasome function. The proteasome is the macromolecular structure responsible for the degradation of misfolded or short-lived proteins in cells, and is composed of four stacked heptameric rings in a barrel-like structure. The long-term goal of this work is to help understand the mechanisms that regulate proteasome assembly in the Mtb system using both mathematical modeling and experimental analyses. The overall objectives in this application are to (1) elucidate the mechanism(s) by which assembly dynamics regulate proteasome formation and (2) determine their role in Mtb proteasome assembly. The central hypothesis is that Mtb proteasome has evolved a set of mechanisms that maximize yield and thus bacterial immune resistance. The rationale for this project is that determination of the mechanisms that regulate Mtb proteasome yield is likely to offer a strong scientific framework whereby new strategies for tuberculosis therapies in patients can be developed. The central hypothesis will be tested by pursuing three specific aims: (1) Evaluate the assembly kinetics of the Mtb proteasome using mathematical and experimental analyses, (2) Develop a biophysical framework to understand interactions between intermediate rings in proteasome assembly and (3) Analyze structures of intermediate rings in Mtb proteasome assembly. In the first aim, a mathematical model will be used to determine the role kinetic parameters play in ring formation and ultimately proteasome assembly. Additionally, Mtb monomers will be used to experimentally measure the kinetics of assembly. For the second aim, a biophysical framework will be developed to study the interactions between monomers and intermediate rings based on their size and structure. Furthermore, a mass-spectrometry approach will be used to identify the size and composition of intermediate rings formed during Mtb proteasome assembly. In the third aim, a cryo-Electron Microscopy approach will be used to analyze the structures of assembly intermediates with atomic resolution. The research proposed in this application is innovative because it focuses on the Mtb proteasome, which has not been sufficiently characterized to date, and because it incorporates both mathematical modeling and experimental methods. The proposed research is significant because it is expected to provide a foundation for the development and future clinical applications of novel Mtb proteasome assembly inhibitors. Ultimately, such knowledge has the potential of offering new opportunities for the development of innovative therapies to treat tuberculosis and other bacterial infections. Moreover, this fellowship is sponsored by Drs. Eric J. Deeds and Joseph A. Loo, who are leaders in their respective fields of computational biology and mass spectrometry. The proposed training plan includes a strong research environment and mentoring team conducive to the applicant’s growth into a highly successful independent researcher.

项目摘要 结核病每年影响多8,000人，结核分枝杆菌 （MTB）可以通过蛋白酶体功能部分抵抗免疫系统。蛋白酶体是 大分子结构负责细胞中错误折叠或短寿命蛋白的降解，并且IS 由四个在类似枪管的结构中的四个堆叠的肝环组成。这项工作的长期目标是帮助 了解使用两种数学的MTB系统中蛋白酶体组装的机制 建模和实验分析。本应用程序中的总体目标是（1）阐明 组装动力学调节蛋白酶体的形成和（2）确定其在MTB中的作用的机制 蛋白酶体组件。中心假设是MTB蛋白酶体已经发展了一组机制 最大化产量，从而使细菌免疫抗性。该项目的理由是确定 调节MTB蛋白酶体产量的机制可能会提供一个强大的科学框架 可以制定患者结核病疗法的策略。中心假设将通过 追求三个具体目标：（1）使用数学来评估MTB蛋白酶体的组装动力学 和实验分析，（2）开发一个生物物理框架来了解之间的相互作用 蛋白酶体组装中的中间环和（3）分析MTB蛋白酶体中间环的结构 集会。在第一个目标中，将使用数学模型来确定动力学参数在 环形成，最终是蛋白酶体组件。此外，MTB单体将用于实验 测量组装的动力学。对于第二个目标，将开发生物物理框架来研究 单体与中环之间的相互作用根据其大小和结构。此外， 质谱法将用于识别形成的中间环的大小和组成 在MTB蛋白酶体组件中。在第三个目标中，将使用冷冻电子显微镜方法 分析具有原子分辨率的组装中间体的结构。这项研究提出了 应用具有创新性，因为它专注于MTB蛋白酶体，但还不够 迄今为止的表征，并且因为它同时结合了数学建模和实验方法。 拟议的研究很重要，因为它有望为开发和 新型MTB蛋白酶体组装抑制剂的未来临床应用。最终，这样的知识具有 提供新的机会来开发创新疗法来治疗结核和 其他细菌感染。此外，该研究金由Drs赞助。 Eric J. Deeds和Joseph A. Loo，谁 是各自计算生物学和质谱领域的领导者。拟议的培训 计划包括强大的研究环境和对申请人成长的导电团队的导电 非常成功的独立研究人员。