Synapt G2-S System with HXMS Automation

描述（由申请人提供）：加州大学圣地亚哥分校的生物分子和蛋白质组学质谱分析设施（BPMSF）希望获得一台带有HXMS自动化的Synapt G2-S系统。该项目的PI首次证明了可以用MALDI质谱法测量氢/氘交换（HX）。她的实验室是最早使用这种实验方法来绘制大分子界面的实验室之一。自1998年以来，BPMSF的MALDI-TOF质谱仪一直用于HDX实验，主要由非质谱专家的生物化学家进行。具有HXMS自动化功能的Synapt G2-S系统是第一个完全自动化的仪器，可以取代MALDI-TOF，供非质谱专家使用。与旧的手动MALDI-TOF方法相比，Waters仪器在灵敏度、自动化和数据分析方面也有许多改进。集成色谱系统基于Waters nanoACQUITY平台，包含一个peltier冷却模块，该模块包含进样和开关阀，在线胃蛋白酶消化和C-18分离柱。该色谱系统与qTOF质谱仪完全集成，该质谱仪结合了双t波，离轴离子转移光学器件，以最高的效率将离子从离子源转移到四极杆质谱分析仪。一套软件协调样品制备，色谱和质谱，并提供快速和自动化的数据原始查看，分析和定量与DynamX软件。主要用户是过去或将来在BPMSF尝试HDX实验的所有研究人员。他们的项目涵盖了美国国立卫生研究院资助的所有研究项目。具有HXMS自动化的Synapt G2-S系统将实现过时的MALDI-TOF方法无法实现的新的重要发现：Komives将测量NF- κ B蛋白抑制剂的“折叠性”以及它们与靶标结合时折叠状态的变化；Ghosh将分析激酶（IKK）的构象变化，IKK磷酸化这些靶向它们降解的抑制剂；诺埃尔将分析烟草5-表马兜铃烯合成酶和其他萜类合成酶的构象变化，他们正在设计这些合成酶来制造新的天然产物；Gianneschi将探索用于药物传递的蛋白质胶束的宏观结构；詹宁斯将分析白细胞介素和激酶的构象变化；Shan将绘制叶绿体信号识别粒子中底物和受体相互作用的图谱；凯将筛选昼夜节律控制蛋白的调制剂。

DESCRIPTION (provided by applicant): The Biomolecular and Proteomics Mass Spectrometry Facility (BPMSF) at the University of California San Diego wishes to acquire a Synapt G2-S System with HXMS Automation. The PI of this project was the first to show that hydrogen/deuterium exchange (HX) could be measured by MALDI mass spectrometry. Her lab was one of the first to use this experimental approach for mapping macromolecular interfaces. Since 1998, the MALDI-TOF mass spectrometer in the BPMSF has been continuously used for HDX experiments, mainly performed by biochemists who were not experts in mass spectrometry. The Synapt G2-S System with HXMS Automation is the first fully-automated instrument that can replace the MALDI-TOF for use by non-mass spectrometry experts. The Waters instrument also provides many improvements in sensitivity, automation, and data analysis compared with the old manual MALDI-TOF approach. The integrated chromatographic system is based on the Waters nanoACQUITY platform and contains a Peltier-cooled module that houses the injection and switching valves, online pepsin digestion and C-18 separation columns. This chromatographic system is fully integrated with the qTOF mass spectrometer which combines dual-T-wave, off-axis ion transfer optics to transfer ions from the ion source to the quadrupole MS analyzer with the highest possible efficiency. A suite of software coordinates the sample preparation, chromatography, and mass spectrometry and provides rapid and automated data raw viewing, analysis and quantitation with the DynamX software. The Major Users are all researchers who have in the past, or will in the future attempt HDX experiments in the BPMSF. Their projects encompass the gamut of NIH-funded research. The Synapt G2-S System with HXMS Automation will enable new and important discoveries not possible with the outdated MALDI-TOF approach: Komives will measure the "foldedness" of inhibitors of NF- kappa B proteins and changes in their folded state upon binding to their targets; Ghosh will analyze conformational changes in the kinase (IKK) that phosphorylates these inhibitors targeting them for degradation; Noel will analyze conformational changes in Tobacco 5-epi-aristolochene synthase and other terpene synthases which they are engineering to make novel natural products; Gianneschi will probe the macrostructure of proteinaceous micelles for drug delivery; Jennings will analyze the conformational changes in interleukins and kinases; Shan will map the substrate- and receptor- interactions in the chloroplast Signal Recognition Particle; and Kay will screen modulators of circadian rhythm-control proteins.