Native Mass Spectrometry Guided Structural Biology Center

天然质谱引导结构生物学中心

• 批准号: 10629935
• 负责人: Vicki H. Wysocki
• 金额: $ 135.1万
• 依托单位: OHIO STATE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-09-19 至 2028-07-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10629935
• 关键词: Acceleration; Agreement; Antibodies; Architecture; Area; Automobile Driving; Binding; Biomedical Technology; Blood capillaries; Buffers; Capillary Electrophoresis; Charge; Collaborations; Communication; Communities; Complement; Complex; Computer Models; Computing Methodologies; Coupled; Couples; Coupling; Data; Detection; Development; Devices; Disease; Dissociation; Educational workshop; Electrospray Ionization; Fostering; Fourier Transform; Funding; Future; Genetic; Geography; Goals; Grant; HIV; Heterogeneity; Human Resources; Industrialization; Institution; Ions; Journals; Laboratories; Leadership; Legal patent; Licensing; Lipids; Macromolecular Complexes; Malignant Neoplasms; Mass Spectrum Analysis; Measurement; Membrane Glycoproteins; Membrane Proteins; Membrane Structure and Function; Mission; Molecular Conformation; Multiprotein Complexes; Nucleoproteins; Phase; Photometry; Polymorph; Prevention; Procedures; Productivity; Protein Secretion; Proteins; Proteomics; Publications; Quaternary Protein Structure; RNA; Research Personnel; Resolution; Resources; Role; Sampling; Scientific Advances and Accomplishments; Services; Site; Small Business Innovation Research Grant; Specialist; Spectrometry; Structure; Sudden infant death syndrome; Surface; System; Technology; Technology Transfer; Temperature; Thermodynamics; Training; Translating; Translations; Vendor; Viral; Virus Diseases; Water; Work; community engagement; complex data; computerized tools; design; digital; empowerment; flexibility; improved; instrumentation; interest; ion mobility; member; nervous system disorder; non-Native; organizational structure; protein complex; protein oligomer; restraint; stoichiometry; structural biology; tau aggregation; technology development; technology research and development; tool; virtual

We propose conversion of the P41 Resource for Native Mass Spectrometry Guided Structural Biology (2018- ) to an RM1 Biomedical Technology Development and Dissemination (BTDD) Center. The nMSSB Center will produce robust, universally-available, transformative technology for characterizing the assembly and disassembly of macromolecular protein complexes, including the definition of their compositions, PTMs, stoichiometry, structure/architecture/topology, flexibility/stability, and their conformational/binding-induced changes under different conditions. The inclusion of an integrated workflow for native mass spectrometry (nMS) will enable, complement, and expand what is possible with other structural biology (SB) tools. We propose three highly synergistic Technology Development Projects (TDPs) that build from our productive P41 Technology Research and Development. Feasibility for the TDPs has been established through preliminary data and publications in high-quality journals, as well as through the licensing of one version of our SID technology by an MS vendor (Waters Corp.), release of a desalting column (based on our online buffer exchange progress) by another vendor (Thermo) in 2022, and the funding of two SBIR grants (eMSion and Protein Metrics). We continue and expand our relationships with multiple MS vendors/ sustainability partners, biopharma collaborators/industrial advisory board members, and add two technology partners (Ivanov and Coon). Ten geographically and topically/scientifically diverse Driving Biomedical Projects (DBPs) have been selected to drive technology development and extend nMS to leading biomedical laboratories that are tackling e.g., cancer, viral infections, and neurological disorders. Our three TDPs focus on different vendor-neutral tools coupled in multiple ways to maximize future nMS options. TDP1 optimizes a panel of solution-phase separation and measurement tools, including the expansion of two-column online buffer exchange, capillary electrophoresis, and variable temperature electrospray ionization. TDP2 develops improved mass selection using digital quadrupole technology and improved resolution Fourier transform ion mobility (IM) spectrometry, couples surface induced dissociation with multiple IM and MS analyzers including charge detection mass spectrometry, and develops computational methods to use energy-resolved nMS data and IM to identify “near-native” computational models. TDP3 develops collision- and surface-induced unfolding to screen biomolecular stability, adapts TDP technologies for vendor-specific complex down/native-omics approaches, and constructs integrated computations that utilize synergistic MS restraints to predict protein quaternary structures. Our administration & management structure provides leadership, coordinates communication/collaboration, and drives sustainability. Our community engagement plan continues dissemination and training to include publications, technology transfer to beta testers and vendors, an annual nMS short course, virtual and hands-on workshops, and multiple “sustainability sites” where nMS workflows will be integrated with other SB tools for non-expert and expert users.

我们建议将P41资源转换为原生质谱指导的结构生物学（2018-） RM 1生物医学技术开发和传播中心（BTDD）。nMS的CSB中心将 产生强大的、普遍可用的、变革性的技术来表征组件， 大分子蛋白质复合物的分解，包括其组成的定义，PTM， 化学计量、结构/架构/拓扑、柔性/稳定性以及它们的构象/结合诱导 在不同的条件下变化。包括用于天然质谱（nMS）的集成工作流程 将启用，补充和扩展其他结构生物学（SB）工具的可能性。我们提出了三 高度协同的技术开发项目（TDPs），以我们的生产性P41技术为基础 研发的通过初步数据确定了技术发展方案的可行性， 在高质量的期刊上发表，以及通过授权我们的SID技术的一个版本， MS供应商（沃茨Corp.），发布一个缓冲区列（基于我们的在线缓冲区交换进度）， 另一家供应商（Thermo）在2022年，以及两个SBIR赠款（eMSion和Protein Side）的资金。我们继续 并扩大我们与多个MS供应商/可持续发展合作伙伴、生物制药 合作者/工业顾问委员会成员，并增加了两个技术合作伙伴（伊万诺夫和库恩）。十 选择了地理和主题/科学多样的生物医学项目（DBP）来推动 技术开发并将nMS扩展到领先的生物医学实验室，癌症，病毒性的 感染和神经系统疾病。我们的三个TDP侧重于不同的供应商中立的工具， 最大化未来NMS选项的方法。TDP 1优化了溶液相分离和测量面板 工具，包括扩展双柱在线缓冲液交换，毛细管电泳和可变 温度电喷雾电离。TDP 2使用数字四极杆开发改进的质量选择 技术和改进的分辨率傅里叶变换离子迁移率（IM）光谱，耦合表面诱导 解离与多个IM和MS分析仪，包括电荷检测质谱，并开发 计算方法使用能量分辨的nMS数据和IM来识别“近原生”计算模型。 TDP 3开发碰撞和表面诱导的解折叠以筛选生物分子稳定性， 用于供应商特定的复杂down/native-omics方法的技术， 利用协同MS约束来预测蛋白质四级结构的计算。我们的行政及 管理结构提供领导，协调沟通/协作，并推动可持续发展。 我们的社区参与计划继续传播和培训，包括出版物、技术 向beta测试人员和供应商转移，每年的nMS短期课程，虚拟和实践研讨会，以及多个 “可持续性网站”，其中nMS工作流程将与非专家和专家用户的其他SB工具集成。