Native Mass Spectrometry Guided Structural Biology Center

我们建议将P41资源转换为天然质谱学指导的结构生物学(2018-) RM1生物医学技术开发和传播(BTDD)中心。网管SB中心将 生产强大的、普遍可用的、变革性的技术，以表征组件和 大分子蛋白质复合体的拆解，包括其组成的定义，PTMS， 化学计量学、结构/架构/拓扑、灵活性/稳定性及其构象/结合诱导 在不同的条件下会发生变化。包含本地质谱学(NMS)的集成工作流程 将启用、补充和扩展其他结构生物学(SB)工具所能实现的功能。我们提出三个建议 从我们的生产性P41技术构建的高度协同的技术开发项目(TDP) 研究和开发。全港发展计划的可行性已透过初步数据及 在高质量期刊上发表文章，以及通过由 MS供应商(Waters Corp.)，发布脱盐柱(基于我们的在线缓冲区交换进度)，由 另一家供应商(Thermo)在2022年，并资助了两个SBIR赠款(eMSion和蛋白质指标)。我们继续 并扩展我们与多家MS供应商/可持续发展合作伙伴、生物制药 合作者/产业顾问委员会成员，并增加两个技术合作伙伴(Ivanov和Coon)。十 已经选择了地理和局部/科学上多样化的推动生物医学项目(DBP)来推动 技术开发并将NMS扩展到领先的生物医学实验室，这些实验室正在研究癌症、病毒等 感染和神经紊乱。我们的三个TDP专注于不同的供应商中立工具，这些工具结合在多个 最大化未来网管选项的方法。TDp1优化了一组溶液相分离和测量 工具，包括扩展的双柱在线缓冲液交换、毛细管电泳法和变量 温度电喷雾电离。TDP2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，电偶表面诱导 与包括电荷检测质谱仪在内的多个IM和MS分析器分离，并开发 使用能量分辨的NMS数据和IM来识别“接近本地的”计算模型的计算方法。 TDP3开发碰撞和表面诱导的去折叠来筛选生物分子稳定性，适应TDP 供应商特定的复杂羽绒/原生组学方法的技术，以及集成的结构 利用协同MS约束来预测蛋白质四级结构的计算。我们的政府和 管理结构提供领导力、协调沟通/协作并推动可持续性。 我们的社区参与计划继续进行传播和培训，以包括出版物、技术 转到测试员和供应商、一年一度的NMS短期课程、虚拟和实践研讨会以及多个 “可持续性站点”，NMS工作流程将与其他SB工具集成，供非专家和专家用户使用。

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.