High throughput X-ray footprinting mass spectrometry (XFMS)

高通量 X 射线足迹质谱 (XFMS)

• 批准号: 10708045
• 负责人: CORIE Y RALSTON
• 金额: $ 14.25万
• 依托单位: UNIVERSITY OF CALIF-LAWRENC BERKELEY LAB
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-09-01 至 2027-08-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10708045
• 关键词: Automation; Award; Binding; Biological; Buffers; Chemistry; Collection; Communities; Complex Mixtures; Computer software; Crystallography; Data; Data Analyses; Data Collection; Databases; Deuterium; Development; Diameter; Digestion; Dose; Exclusion; Exposure to; Fingerprint; Fluorescence; Fluorescence Spectroscopy; Grant; Hour; Hybrids; Hydrogen; Hydroxyl Radical; In Situ; Individual; Investigation; Kinetics; Label; Laboratories; Learning; Liquid Chromatography; Liquid substance; Maps; Mass Spectrum Analysis; Methods; Molecular Conformation; Molecular Sieve Chromatography; National Institute of General Medical Sciences; Nucleic Acids; Positioning Attribute; Process; Proteins; Protocols documentation; Pump; Raman Spectrum Analysis; Reporting; Resources; Roentgen Rays; Sampling; Shotguns; Signal Transduction; Site; Solvents; Spectrum Analysis; Speed; Structure; Syringes; System; Technology; Time; Tube; X ray spectroscopy; base; beamline; biological systems; chromophore; data access; experimental study; flexibility; instrument; instrumentation; liquid chromatography mass spectrometry; macromolecular assembly; macromolecule; operation; protein complex; protein structure; sample collection; spectroscopic data; structural biology; success; time use; tool; web site

Technology Operations Core 4: Abstract Technology Operations Core 4 (TOC4) integrates the method of X-ray footprinting mass spectrometry (XFMS) into the ALS-ENABLE resource, providing resource users with a solution state structural biology method that is highly complementary to macromolecular crystallography (MX, TOCs1&3) and small angle X-ray scattering (SAXS, TOC2). XFMS is an in situ hydroxyl radical (•OH) labeling method based on well-established solvent accessibility protocols similar to Fenton chemistry and hydrogen deuterium exchange, sharing the same liquid chromatography-mass spectrometry (LCMS) data collection platform. The method provides residue-level structural information on proteins and/or nucleic acid in the solution state, and can be used for time-resolved studies and on complex mixtures of proteins in solution. Because of its versatility, the method has been especially valuable in obtaining structural and kinetics information that is highly complementary to the information obtained using the other more well-known structural biology methods such as MX an SAXS. The XFMS method has a vibrant, growing user base, has advanced substantially over the previous decade in automation and throughput, and is now close to the same level of routine use as MX and SAXS. In the last four years in particular, the instrumentation developed as part of a current NIGMS R01 award has significantly advanced the throughout and accessibility of the method for the larger biomedical community, including the development of an automated liquid sample delivery system, a remote-accessible beamline control interface, and automated LCMS data analysis. Through TOC4, we will make available our automated, high throughput, and hybrid spectroscopy- XFMS platform to ALS-ENABLE users through remote access, onsite and mail-in options. We will integrate information about XFMS and access to databases through the ALS-ENABLE website (als-enable.lbl.gov) so that users will have a single front-facing portal through which to learn about and access data from all the structural biology beamlines at the ALS. The integration of these technologies within the ALS-ENABLE resource will extend the structural biology capabilities at the ALS such that even more challenging biological systems can be studied, thus providing the national biomedical community with a powerful and increasingly versatile new tool for investigation of macromolecular assemblies.

技术操作核心4：摘要 技术操作核心4（TOC4）整合了X射线足迹质谱法（XFM）的方法 进入ALS启用资源，为资源用户提供解决方案状态结构生物学方法，这是 高分分子晶体学（MX，TOCS1和3）和小角度X射线散射 （SAXS，TOC2）。 XFMS是基于公认溶剂的原位羟基自由基（•OH）标记方法 可访问性方案类似于芬顿化学和氢氘交换，共享相同的液体 色谱 - 质谱法（LCMS）数据收集平台。该方法提供居住级别 溶液状态下蛋白质和/或核酸的结构信息，可用于时间分辨 研究和溶液中蛋白质的复杂混合物。由于其多功能性，该方法尤其是 获得高度完整的结构和动力学信息的价值 使用其他更知名的结构生物学方法，例如MX和SAX。 XFMS方法具有 充满活力的，增长的用户群在过去十年中的自动化和吞吐量中取得了长足的进步， 现在，与MX和SAXS相同的常规使用级别。特别是在过去的四年中， 作为当前NIGMS R01奖的一部分而开发的仪器在整个过程中都显着推进了 该方法可访问大型生物医学界的可及性，包括开发自动化的 液体样品输送系统，可远程访问的光束线控制接口和自动LCMS数据 分析。通过TOC4，我们将提供自动化，高通量和杂种光谱法 - XFMS平台通过远程访问，现场和邮寄选项向ALS-Enable用户提供。我们将整合 有关XFM的信息以及通过ALS-Enable网站（Als-enable.lbl.gov）访问数据库的信息，以便 用户将拥有一个单一的面向门户网站，可以通过该门户从所有结构中学习和访问数据 ALS的生物束线。这些技术在ALS-Enable资源中的集成将 扩展ALS的结构生物学能力，使得更具挑战性的生物系统可以是 研究菌 大分子组件的研究。