Electron-Transfer Dissociation Tandem Mass Spectrometry

描述(申请人提供)：本提案的主要目标是开发一种新型的MS接口设备，能够在任何质谱仪上提供高效的ECD/ETD类型的多肽和蛋白质的裂解，包括低成本的台式仪器，如单四极杆和三四极杆。在这项建议中，我们建议使用在第一阶段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方法特有的各种优点。赫德技术的好处是显而易见的，也是实质性的。HRID技术可以用作与标准CAD碎片化方法正交的方法，以最大限度地减少假阳性的概率，这被认为是“自下而上”识别策略中的严重问题之一。重要的是，HRID技术能够提供ECD/ETD类型的串联MS数据，即使对于单电荷肽来说，这肯定是非常吸引人的特征，例如，对于AP-MALDI用户。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)大的完整蛋白质的有效测序和(2)直接确定蛋白质的翻译后修饰位置。 公共卫生相关性：蛋白质组学工具的应用在现代基础科学、药物开发和临床应用中发挥着重要作用。我们提出了一种利用串联质谱仪进行蛋白质鉴定的新技术，这是当今蛋白质组学中的一种基本策略。

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.