Electrospray Ionization (ESI) mass spectrometer with liquid chromatography for proteome analysis

该应用程序用于电喷雾电离质谱仪，包括具有出色灵敏度、分辨率和质量准确度的色谱，能够进行不同类型的扫描。这包括不同的碎片化、数据依赖性和数据独立性采集、质量窗口扫描、靶向分析以及使用无标记和同位素标记方法的稳健定量。该设备主要用于“植物生物化学”部门的研究，该部门现有的质谱仪已经过时。主要研究方向为叶绿体质量控制系统和叶绿体生物发生的泛素-蛋白酶体调控、不同质体类型分化的蛋白质组分析以及叶绿体功能的磷酸化调控。在所有情况下，需要高灵敏度的蛋白质组分析，例如，以表征叶绿体质量控制系统的组成部分的突变体，并了解其不同功能的串扰。由于这些过程在转录后发生作用，因此必须在蛋白质组水平上进行分析。这同样适用于质体的分化，特别是在分析蛋白质输入系统的功能时，以及分析翻译后修饰及其功能时。该装置应适合于鉴定非常低丰度的肽的翻译后修饰并对其进行稳健定量。分析深度必须很高，因为例如胞质溶胶中质体前体蛋白的N-末端肽经历高周转，并且除了它们已经非常低的丰度之外，还非常不稳定。该设备将在未来的设施中安装，大约30%的时间将用于委托工作。这意味着它必须坚固耐用，相对易于操作，并且维护要求低。

This application is for an electrospray ionization mass spectrometer including chromatography with excellent sensitivity, resolution and mass accuracy, which is able to carry out different types of scans. This includes different fragmentations, data-dependent and data-independent acquisitions, mass window scans, targeted analyses and robust quantifications using label-free and isotope-labeled methods. This device is primarily intended to be used for research at the “Plant Biochemistry” department, where the existing mass spectrometers are out of date. Research topics are the chloroplast quality control system and regulation of chloroplast biogenesis via the ubiquitin-proteasome system, proteome analyses of different plastid type differentiation and the operational control of chloroplast functions via phosphorylation. In all cases, highly sensitive proteome analysis is required, e.g. to characterize mutants of components of the chloroplast quality control system and to understand the crosstalk of its different functions. Since these processes take effect post-transcriptionally, the analyses must be carried out at the proteome level. The same applies to the differentiation of plastids, especially when it comes to analyze the functionality of the protein import system, and to the analysis of post-translational modifications and their function. The device should be suitable to identify post-translational modifications of very low abundant peptides and to quantify them robustly. Analytical depth must be high, because e.g. the N-terminal peptides of plastid precursor proteins in the cytosol are subject to a high turnover and, in addition to their already very low abundance, are also highly unstable. The device is to be set up in a facility in the future, and around 30% of it’s time will be made available for commissioned work. This means that it has to be robust, relatively easy to operate and have low maintenance requirements.