Resource for Native Mass Spectrometry Guided Structural Biology

天然质谱引导结构生物学资源

• 批准号: 9978835
• 负责人: Vicki H. Wysocki
• 金额: $ 122.94万
• 依托单位: OHIO STATE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-07-01 至 2023-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9978835
• 关键词: Address; Affinity; American; Architecture; Area; Automobile Driving; Beds; Behavior; Binding; Biological; Biomedical Research; Capillary Electrophoresis; Cataract; Cell physiology; Collaborations; Communities; Complement; Complex; Computer software; Coupled; Cryoelectron Microscopy; Crystallization; DNA; Data; Defect; Devices; Digit structure; Disease; Dissociation; Educational workshop; Exclusion; Foundations; Functional disorder; Generations; Goals; HIV; HIV-1; Heterogeneity; Human Resources; Individual; Institution; International; Ion-Exchange Chromatography Procedure; Label; Laboratories; Lead; Letters; Lipids; Macromolecular Complexes; Mass Spectrum Analysis; Measurement; Membrane Proteins; Methods; Mission; Modeling; Molecular Conformation; Neurologic; Pathology; Pattern; Physiological Processes; Physiology; Prevention; Procedures; Protein Engineering; Proteins; Proteomics; RNA; RNA Binding; RNase P; Regulation; Research Personnel; Resolution; Resources; Roentgen Rays; Role; Sampling; Science; Scientific Advances and Accomplishments; Scientist; Services; Site; Societies; Structure; Surface; Technology; Technology Transfer; Testing; Texas; Time; Training; United States; Universities; Vendor; Viral Hemorrhagic Fevers; Viral Oncogene Proteins; Viral Packaging; Water; adduct; austin; base; biomacromolecule; clinically significant; computational chemistry; computerized tools; design; experimental study; genomic RNA; innovation; instrument; instrumentation; interest; ion mobility; ionization; macromolecular assembly; nervous system disorder; novel; oligomycin sensitivity-conferring protein; outreach; protein complex; restraint; spatiotemporal; stoichiometry; structural biology; technology development; technology research and development; tool; transmission process

We propose a Resource for Native Mass Spectrometry Guided Structural Biology that will be led by a team of scientists including experts in MS instrumentation (Wysocki, OSU; Russell, Texas A&M), separation science coupled to ionization (Olesik, OSU; Badu, OSU; Holland; WVU), and computational chemistry (Lindert, OSU). The goal of this Resource is to build and validate an integrated workflow for structural characterization of protein:protein, membrane protein:lipid, and protein:RNA complexes that are critical for an array of cellular and organismal processes. There is a growing appreciation of the pivotal role and utility of MS-based approaches for structural characterization of biomacromolecules, filling critical gaps and complementing other structural biology tools. Thus, our workplan leverages innovative MS methods to determine: (i) m/z of all binding partners and the intact complex, (ii) component stoichiometry, (iii) heterogeneity, if present, (iv) the relative topology/architecture/ conformational diversity of components in the complex, and (v) the hierarchy of assembly, including binding affinities of individual subunits. Investigators from across the United States (in WA, OR, UT, CA, AZ, TX, TN, OH, MD, MA) and international sites will contribute challenging biomedical projects on topics including viral hemorrhagic fevers, HIV, cataract formation, and neurological disorders. Our workflow is designed to advance understanding of: (i) how formation of and dynamic changes in wide- ranging macro-molecular assemblies determine their biological roles, (ii) how alterations in assembly/architecture of these complexes lead to disease, and (iii) foundational principles for building synthetic mimics needed for biomedical applications. In this Resource, ten Driving Biomedical Projects (DBPs) provide biomedical structural characterization challenges and serve as drivers and test beds for five Technology Research and Development (TR&D) projects. The TR&Ds provide: (i) effective separation methods to purify and deliver native macromolecular complexes well suited for MS, (ii) effective surface-induced dissociation and UV-photodissociation technologies, (iii) measurement of the intact complexes and their non-covalent (sub-complex) and covalent dissociation products with high resolution ion mobility (IM) and/or MS, and (iv) computational tools for structure prediction. Computational tools will use restraints from surface-induced dissociation patterns and collision cross sections from IM-MS experiments, and from MS-based solution measurements (H/D exchange and covalent labeling). Existing ties between the TR&Ds and instrument companies (Waters, Thermo, Bruker, Sciex, Phenomenex) as well as a national laboratory (PNNL) will aid technology development and expedite technology dissemination. Our training and dissemination activities (e.g., workshops, beta device installations) are designed to increase outreach and maximize Resource payoffs.

我们提出了一个本地质谱引导结构生物学资源，该资源将由 由包括 MS 仪器专家在内的科学家团队（俄勒冈州立大学 Wysocki；德克萨斯 A&M 拉塞尔）， 分离科学与电离耦合（Olesik，OSU；Badu，OSU；Holland；WVU）和计算 化学（林德特，俄勒冈州立大学）。该资源的目标是构建和验证集成的工作流程 蛋白质：蛋白质、膜蛋白：脂质和蛋白质：RNA 复合物的结构表征 对于一系列细胞和有机体过程至关重要。人们越来越欣赏 基于 MS 的生物大分子结构表征方法的关键作用和实用性， 填补关键空白并补充其他结构生物学工具。因此，我们的工作计划利用 创新的 MS 方法可确定：(i) 所有结合伴侣和完整复合物的 m/z，(ii) 成分化学计量，(iii) 异质性（如果存在），(iv) 相对拓扑/结构/ 复合物中组分的构象多样性，以及（v）组装的层次结构，包括 各个亚基的结合亲和力。来自美国各地（华盛顿州、俄勒冈州、犹他州、 加利福尼亚州、亚利桑那州、德克萨斯州、田纳西州、俄亥俄州、马里兰州、马里兰州）和国际站点将贡献具有挑战性的生物医学项目 主题包括病毒性出血热、艾滋病毒、白内障形成和神经系统疾病。我们的 工作流程旨在增进对以下方面的理解：（i）广泛的信息如何形成和动态变化。 范围广泛的大分子组装体决定了它们的生物学作用，(ii) 这些综合体的组装/建筑会导致疾病，以及（iii）建筑的基本原则 生物医学应用所需的合成模拟物。在本资源中，十个推动生物医学项目 （DBP）提供生物医学结构表征挑战并充当驱动程序和测试台 五个技术研发 (TR&D) 项目。 TR&D 提供：(i) 有效 纯化和提供非常适合 MS 的天然大分子复合物的分离方法，(ii) 有效的表面诱导解离和紫外光解离技术，(iii) 的测量 完整的复合物及其非共价（亚复合物）和共价解离产物具有高 分辨率离子淌度 (IM) 和/或 MS，以及 (iv) 用于结构预测的计算工具。 计算工具将使用表面诱导解离模式和碰撞交叉的限制 来自 IM-MS 实验和基于 MS 的溶液测量（H/D 交换和 共价标记）。 TR&D 和仪器公司（Waters、Thermo、 Bruker、Sciex、Phenomenex）以及国家实验室 (PNNL) 将协助技术开发 并加快技术传播。我们的培训和传播活动（例如研讨会、测试版 设备安装）旨在扩大覆盖范围并最大化资源回报。