High throughput X-ray footprinting mass spectrometry (XFMS)

技术运营核心4：摘要 技术操作核心4（TOC 4）集成了X射线足迹质谱法（XFMS） ALS-ENABLE资源，为资源用户提供解决方案状态结构生物学方法， 与大分子晶体学（MX，TOC 1&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 与芬顿化学和氢氘交换相似的可访问性协议，共享相同的液体 色谱-质谱（LCMS）数据收集平台。该方法提供了残留水平， 在溶液状态下的蛋白质和/或核酸的结构信息，并且可以用于时间分辨 研究和复杂的蛋白质混合物在溶液中。由于其多功能性，该方法特别适用于 在获得与所获得的信息高度互补的结构和动力学信息方面有价值 使用其他更知名的结构生物学方法，如MX和SAXS。XFMS方法具有 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长足的进步， 并且现在接近于与MX和SAXS相同的常规使用水平。特别是在过去的四年里， 作为当前NIGMS R 01奖项的一部分而开发的仪器大大提高了整个系统的性能， 更大的生物医学界的方法的可访问性，包括开发自动化的 液体样品输送系统、远程可访问的光束线控制接口和自动化LCMS数据 分析.通过TOC 4，我们将提供我们的自动化，高通量和混合光谱- XFMS平台通过远程访问、现场和邮寄选项向ALS启用用户提供。我们将整合 通过ALS-ENABLE网站（als-enable.lbl.gov）提供有关XFMS的信息和对数据库的访问， 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化 ALS的生物光束线这些技术在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，从而为国家生物医学界提供了一个强大的，越来越多功能的新工具， 大分子组装研究。

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.