Gas-phase Bio-conjugation in the Tandem Mass Spectrometry of Peptides Proteins

肽蛋白串联质谱中的气相生物共轭

• 批准号: 10333337
• 负责人: SCOTT A MCLUCKEY
• 金额: $ 32.71万
• 依托单位: PURDUE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 1991
• 资助国家: 美国
• 起止时间: 1991-09-01 至 2024-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10333337
• 关键词: Area; Biological; Biomedical Research; Charge; Chemicals; Chemistry; Communities; Complex; Development; Digestion; Enzymes; Gases; Health; Ions; Literature; Mass Spectrum Analysis; Measurement; Methodology; Methods; Molecular Biology; Peptides; Phase; Play; Post-Translational Protein Processing; Proteins; Proteomics; Reaction; Research; Role; Sampling; Science; Virus; Work; base; experimental study; functional group; improved; lipidomics; metabolomics; millisecond; novel; novel strategies; protein complex; tandem mass spectrometry

The overall thrust of this research initiative is to develop new and improved means for the structural characterization of ions of biomedical relevance, with particular emphasis on those formed from peptides derived from enzymatic or chemical digestion, whole proteins, and protein complexes. The objective is to develop a new direction in tandem mass spectrometry that will enable a wide range of novel strategies to deal with biomolecule identification/characterization problems. The vast majority of tandem mass spectrometry experiments rely on fragmentation as the key structurally informative reaction. In this work, we seek to add the capability for selective covalent or non-covalent derivatization of gaseous ions within the context of an MSn experiment. The development of specific gas-phase chemistries fundamentally enables the implementation of a very wide range of novel applications. Just as bio-conjugation chemistries in solution have become broadly used by the molecular biology research community in a wide range of measurement strategies, the ability to effect gas-phase bio-conjugation will enable many novel approaches to structural characterization. Among the interesting features of the gas-phase approach are that it is fast (i.e., tens of milliseconds timescale) and it completely avoids sample manipulation issues. The work will involve the development of novel selective reactions, many of which will be inspired by the rich bio-conjugation literature, the development of peptide ion-based applications, and the development of whole protein/protein complex ion-based applications, as reflected in the specific aims: Specific Aim 1: Develop top-down protein characterization methods based on selective "enzyme-like" fragmentation enabled by the functional group specific ion/ion reactions discovered in the early years of this project. Specific Aim 2: Develop applications for the use of selective ion/ion reactions in the Msn of peptide and protein ions for the identification/characterization of sub-stoichiometric post-translational modifications in both bottom-up and top-down proteomics scenarios. Specific Aim 3: Develop novel approaches for protein and protein complex characterization using ion/ion charge transfer strategies. These approaches will be applied to protein complexes and large biological complexes (e.g., viruses).

这项研究的总体目标是开发新的和改进的手段， 生物医学相关离子的表征，特别强调由肽形成的离子 来源于酶或化学消化、完整蛋白质和蛋白质复合物。目标是 发展串联质谱的新方向，使广泛的新策略能够处理 具有生物分子识别/表征问题。绝大多数串联质谱 实验依赖于片段化作为关键的结构信息反应。在这项工作中，我们试图增加 在MSn环境中选择性共价或非共价衍生气态离子的能力 实验特定气相化学的发展从根本上实现了 of a very wide宽range范围of novel新applications应用.正如溶液中的生物共轭化学已经成为 被分子生物学研究界广泛用于广泛的测量策略， 实现气相生物共轭的能力将使许多新的结构表征方法成为可能。 气相方法的有趣特征之一是它是快速的（即，几十毫秒的时间尺度） 并且完全避免了样品操作问题。这项工作将涉及小说的发展 选择性反应，其中许多将受到丰富的生物缀合文献的启发， 基于肽离子的应用，以及基于全蛋白质/蛋白质复合物离子的应用的开发， 具体目标体现在： 具体目标1：开发基于选择性“酶样”的自上而下的蛋白质表征方法 在本发明的早期发现的由官能团特异性离子/离子反应实现的碎片化 项目 具体目标2：开发在肽的MSN中使用选择性离子/离子反应的应用， 蛋白质离子，用于鉴定/表征亚化学计量的翻译后修饰， 自下而上和自上而下的蛋白质组学方案。 具体目标3：开发使用离子/离子的蛋白质和蛋白质复合物表征的新方法 电荷转移策略这些方法将应用于蛋白质复合物和大型生物 复合物（例如，病毒）。