MICROMASS HYBRID QUADRUPOLE--TOF MASS SPECTROMETER

这项申请的主要目的是购买一台混合四极正交飞行时间质谱仪(Q-TOF)。UAB研究人员开展的研究工作越来越多地是鉴定参与特定细胞功能(信号、细胞形状、细胞运动等)的生物化学或化学修饰形式的蛋白质。在单个细胞系和患病组织中。Q-TOF灵敏度的提高将允许对蛋白质的内源性翻译后修饰进行检测。化学修饰可以是有意的(研究结构或机制假说的探针)，也可以是结果的(治疗药物、其代谢物之一或其他外源物质的作用)。这些实验将包括氢-氚交换实验，从而能够研究蛋白质折叠和蛋白质-蛋白质相互作用。在许多情况下，可供分析的蛋白质和/或其蛋白水解肽的数量受到严格限制(中飞摩尔范围)，比质谱学共享设施中现有的质谱仪或任何化学测序方法可以检测到的数量少10-100倍。Q-TOF仪器将四极离子选择与离子碎裂和飞行时间分析仪相结合，为解决这些问题提供了必要的灵敏度提高。在相关的应用中，研究人员将利用Q-TOF的更宽的m/z范围(0-8000)来研究蛋白质分子离子的低电荷态。这可能是由于(1)特定蛋白质的氨基酸组成，92)在神经pH下进行电喷雾实验，或(3)在研究蛋白质-蛋白质和蛋白质-DNA结合过程中。总而言之，微质量Q-TOF质谱仪将大大增强UAB广大教职员工进行我们NIH资助的生物医学研究的能力。

The principal purpose of this application is the purchase of a hybrid quadrupole-orthogonal time-of-flight mass spectrometer (Q-tof). An increasing part of th research effort conducted by UAB investigators is identification of the biochemically or chemically modified forms of proteins mediating specific cellular functions (signalling, cell shape, cell motility, etc.) in individual cell lines and in diseased tissues. The increased sensitivity of the Q-tof will allow endogenous, post- translational modifications of proteins to be examined. Chemical modifications may be deliberate (probes to investigate structural or mechanistic hypotheses) or consequential (in the action of a therapeutic drug, one of its metabolites, or other xenobiotics). These will include hydrogen-deuterium exchange experiments which enable the study of protein folding and protein-protein interactions. In many cases, the amounts of the proteins and/or their proteolytic peptides available for analysis is strictly limited (mid-femtomole range), 10-100 times less than can be detected by the existing mass spectrometers in the Mass Spectrometry Shared Facility, or by any chemical sequencing method. The Q-tof instrument, by combining quadrupole ion selection with ion fragmentation and a time-of-flight analyzer, provides the necessary increase in sensitivity to attack these problems. In related applications, investigators will take advantage of the wider m/z range (0-8,000) of the Q-tof to study lower charge states of molecular ions of proteins. This may be a consequence of (1) the amino acid composition of a particular protein, 92) carrying out electrospray experiments at neural pH, or (3) during the study of protein-protein and protein-DNA binding. In summary, the Micromass Q-tof mass spectrometer will substantially enhance the capability of a wide range of UAB faculty carrying our NIH-funded biomedical research.