Electron-Transfer Dissociation Tandem Mass Spectrometry

描述（由申请人提供）：本提案的主要目标是开发一种新型的质谱接口设备，能够在任何质谱仪上提供高效的ECD/ etd型肽和蛋白质碎片，包括低成本的台式仪器，如单和三四极杆。在本提案中，我们建议使用在I期研究中开发的新型羟基自由基诱导解离（HRID）技术，该技术具有ECD/ETD方法特有的各种优势。HRID技术的好处是显而易见的。HRID技术可以作为标准CAD碎片化方法的一种正交方法，以最大限度地减少假阳性的概率，而假阳性是“自下而上”识别策略中的一个严重问题。重要的是，HRID技术能够提供ECD/ etd型串联质谱数据，甚至可以用于单电荷肽，这对于AP-MALDI用户来说无疑是非常有吸引力的功能。我们相信，与HRID技术一起使用的“自上而下”方法将提供与ECD和ETD方法相同的关键特征，即：(1)对大型完整蛋白质的高效测序；(2)直接确定蛋白质的翻译后修饰位点。

DESCRIPTION (provided by applicant): The main goal of this proposal is to develop a novel MS interface device, capable to provide an efficient ECD/ETD-type fragmentation of peptide and proteins at any mass spectrometer including low-cost bench-top instruments such as single- and triple- quadrupoles. In this proposal we suggest using the novel hydroxyl radical induced dissociation (HRID) technique, developed in Phase I Studies, that offers various advantages, specific to ECD/ETD methods. The benefits of the HRID technology are clear and substantial. The HRID technique can be used as an orthogonal to standard CAD fragmentation method in order to minimize the probability of false positives which are considered as one of the serious problems in the "bottom-up" identification strategy. Importantly, the HRID technique is able to provide ECD/ETD-type tandem MS data even for singly charged peptides that will certainly be very attractive feature, for instance, for AP-MALDI users. We believe that the "top-down" approach used with the HRID technique will provide the same key features as ECD and ETD methods, namely: (1) the efficient sequencing of large intact proteins and (2) the direct determination of the site of post-translational modifications in proteins. PUBLIC HEALTH RELEVANCE: The application of proteomics tools plays an important role in modern basic science, drug discovery and clinical applications. We propose a new technology for protein identification using tandem mass spectrometry, an essential strategy in proteomics today.