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仪(BPMSF)希望获得具有HXMS自动化的Synapt G2-S系统。该项目的等电点首次表明可以用MALDI质谱仪测量氢/氢交换(HX)。她的实验室是第一批使用这种实验方法绘制大分子界面图的实验室之一。自1998年以来，BPMSF中的MALDI-TOF质谱计一直用于HDX实验，主要由不是质谱学专家的生物化学家进行。采用HXMS Automation的Synapt G2-S系统是第一台可取代MALDI-TOF供非质谱学专家使用的全自动化仪器。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。集成的色谱系统基于Waters NanACQUITY平台，包含一个珀尔蒂埃冷却模块，该模块容纳了进样阀和开关阀、在线胃酶消化和C-18分离柱。该色谱系统与qTOF质谱仪完全集成在一起，qTOF质谱仪结合了双T波、离轴离子转移光学元件，以尽可能高的效率将离子从离子源转移到四极杆MS分析器。一套软件协调样品准备、层析和质谱分析，并通过DynamX软件提供快速和自动化的原始数据查看、分析和定量。主要用户都是过去或未来将尝试在BPMSF进行HDX实验的研究人员。他们的项目涵盖了美国国立卫生研究院资助的所有研究。采用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将测量核因子-kappa B蛋白抑制剂的“折叠性”及其与目标结合后折叠状态的变化；Ghosh将分析使这些抑制剂磷酸化以用于降解的激酶(IKK)的构象变化；Noel将分析烟草5-表马兜铃烯合成酶和其他萜烯合成酶的构象变化，他们正在设计这些蛋白质以制造新的天然产品；Gianneschi将探索用于药物输送的胶束的宏观结构；Jennings将分析皮间蛋白和激动酶的构象变化；San将在叶绿体信号识别颗粒中绘制底物与受体的相互作用图；Kay将筛选昼夜节律控制蛋白的调节器。

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.