Resource for Native Mass Spectrometry Guided Structural Biology

天然质谱引导结构生物学资源

• 批准号: 10192745
• 负责人: Vicki H. Wysocki
• 金额: $ 122.92万
• 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/10192745
• 关键词: Address; Affinity; American; Architecture; Area; Automobile Driving; Beds; Behavior; Binding; Biological; Biomedical Research; Capillary Electrophoresis; Cataract; Cell physiology; Collaborations; 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; community engagement; 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.

我们为天然质谱指导的结构生物学提出了一种资源 由一组科学家组成 分离科学耦合到电离（Olesik，OSU； Badu，OSU； Holland; WVU）和计算 化学（Lindert，OSU）。该资源的目的是构建和验证一个集成的工作流程 蛋白质的结构表征：蛋白质，膜蛋白：脂质和蛋白质：RNA络合物，使得 对于一系列细胞和生物过程至关重要。对 基于MS的方法的关键作用和实用性在生物乳清分子的结构表征， 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方法来确定：（i）所有结合伙伴的m/z和完整的复合物，（ii） 组件化学计量学，（iii）异质性，如果存在，（iv）相对拓扑/体系结构/ 综合体中组成部分的构象多样性，以及（v）组件的层次结构，包括 单个亚基的结合亲和力。来自美国各地的调查人员（在华盛顿州或犹他州， CA，AZ，TX，TN，OH，MD，MA）和国际网站将为生物医学项目提供挑战 主题包括病毒出血发烧，HIV，白内障形成和神经系统疾病。我们的 工作流程旨在提高对：（i）广泛的形成和动态变化的理解： 范围宏观分子组件确定其生物学作用，（ii）如何改变 这些综合体的组装/结构导致疾病，以及（iii）建筑的基础原理 生物医学应用所需的合成模拟物。在此资源中，十个驾驶生物医学项目 （DBP）提供生物医学结构表征挑战，并充当驱动因素和测试床 用于五个技术研发（TR＆D）项目。 TR＆D提供：（i）有效 分离方法纯化和传递适合MS的天然大分子复合物，（ii） 有效的表面引起的解离和紫外线解剖技术，（iii）测量 完整的复合物及其非共价（亚复合物）和共价解离产物高 分辨率离子迁移率（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引起的解离模式和碰撞交叉的限制。 IM-MS实验的部分以及基于MS的解决方案测量值（H/D交换和 共价标记）。 TR＆D和仪器公司之间的现有关系（Waters，Thermo， Bruker，Sciex，现象）以及国家实验室（PNNL）将有助于技术开发 并加快技术传播。我们的培训和传播活动（例如，讲习班，Beta 设备安装）旨在增加外展和最大化资源收益。