High throughput X-ray footprinting mass spectrometry (XFMS)

高通量 X 射线足迹质谱 (XFMS)

• 批准号: 10506288
• 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/10506288
• 关键词: Automation; Award; Binding; Biological; Buffers; Caliber; Chemistry; Collection; Communities; Complex Mixtures; Computer software; Crystallography; Data; Data Analyses; Data Collection; Databases; Deuterium; Development; 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; 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.

技术运营核心 4：摘要 技术运营核心 4 (TOC4) 集成了 X 射线足迹质谱 (XFMS) 方法 进入 ALS-ENABLE 资源，为资源用户提供解决状态结构生物学方法，即 与大分子晶体学（MX、TOCs1&3）和小角 X 射线散射高度互补 （SAXS、TOC2）。 XFMS 是一种基于成熟溶剂的原位羟基自由基 (·OH) 标记方法 可访问协议类似于芬顿化学和氢氘交换，共享相同的液体 色谱-质谱（LCMS）数据收集平台。该方法提供残留水平 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，该方法特别受到 对于获得与所获得的信息高度互补的结构和动力学信息很有价值 使用其他更知名的结构生物学方法，例如 MX 和 SAXS。 XFMS 方法有 充满活力、不断增长的用户群，在过去十年中在自动化和吞吐量方面取得了显着进步， 现在已接近 MX 和 SAXS 的日常使用水平。尤其是最近四年， 作为当前 NIGMS R01 奖项的一部分开发的仪器显着推进了整个过程和 该方法对于更大的生物医学界来说是可访问的，包括开发自动化的 液体样品输送系统、远程访问光束线控制接口和自动化 LCMS 数据 分析。通过 TOC4，我们将提供自动化、高通量和混合光谱 - XFMS 平台通过远程访问、现场和邮寄选项为 ALS-ENABLE 用户提供支持。我们将整合 有关 XFMS 的信息以及通过 ALS-ENABLE 网站 (als-enable.lbl.gov) 访问数据库，以便 用户将拥有一个单一的前端门户，通过该门户可以了解和访问来自所有结构的数据。 ALS 的生物学光束线。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，以便可以研究更具挑战性的生物系统 研究，从而为国家生物医学界提供了一个强大且日益通用的新工具 大分子组装体的研究。