INSTRUMENTATION FOR ANALYSIS OF HIGH MASS BIOMOLECULES

高质量生物分子分析仪器

• 批准号: 8265927
• 负责人: SCOTT A MCLUCKEY
• 金额: $ 22.38万
• 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 2014-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8265927
• 关键词: Address; Anions; Biological Markers; Biomedical Research; Cations; Charge; Chemicals; Chemistry; Coupling; Development; Device or Instrument Development; Digestion; Disease; Dissociation; Electron Transport; Gases; Health; Hybrids; Ions; Location; Mass Spectrum Analysis; Measurement; Metals; Methodology; Methods; Modification; Molecular; Nature; Peptides; Phase; Play; Post-Translational Protein Processing; Process; Proteins; Reaction; Reagent; Research; Role; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; Structural Protein; Surface; Time; Work; base; improved; instrument; instrumentation; interest; ion source; ionization; ionization technique; mass spectrometer; meetings; novel; novel strategies; polypeptide; protein complex; research study; tandem mass spectrometry; tool

DESCRIPTION (provided by applicant): The overall thrust of this research initiative is to develop new and improved means for manipulating and interrogating macro-molecular ions with strong emphasis on polypeptide species, including peptides derived from enzymatic or chemical digestion, whole proteins, and protein complexes. The ultimate objective is to maximize the structural information that can be derived from proteins and protein complexes of interest for the purpose of their identification and characterization. The work involves emphases in ion formation, ion chemistry, and instrumentation pertinent to mass spectrometry and tandem mass spectrometry. Gas-phase ions serve as surrogates for the condensed-phase species of interest. Structural information in tandem mass spectrometry is derived both from ion chemistry and the accurate measurement of the masses of the charged reaction products. The nature of the ion (e.g., protonated molecule versus radical cation) plays a major role in its chemistry, as do the reaction conditions. This research is based on the premise that new ion chemistries, new tools, and new methodologies can address many of the current limitations in the ability of mass spectrometry to derive structural information from large polypeptide species. It is also based on the premise that it is both desirable and possible to separate the ionization process from the ion manipulation/interrogation and mass analysis steps so that each can be optimized individually. To further increase the protein structural information accessible from mass spectrometry and tandem mass spectrometry, we propose the following specific aims: Specific Aim 1: New and improved ion chemistry-based approaches for protein identification and structural characterization. Specific Aim 2: New instrumentation and methodologies for tandem mass spectrometry of peptides and proteins in "hybrid" tandem mass spectrometers. Specific Aim 3: Development of novel approaches for tandem mass spectrometry of proteins ionized via matrix-assisted laser desorption ionization (MALDI). These aims will be met by the development of novel ion/ion reactions for ion transformation, an expanded range of ion activation methods, instrument development to support the new ion chemistries, and the development of new approaches to protein identification/characterization that are enabled by the advances in ion chemistry and instrumentation. PUBLIC HEALTH RELEVANCE: This project is directed at improved protein identification/characterization. The ability to identify a protein and to characterize its post-translational modifications (i.e., identities and locations of modifications) underlie much of fundamental biomedical research. This capability is also key to the identification of protein biomarkers and their correlation with disease. This project seeks to address this task by greatly expanding the information content accessible via mass spectrometry.

描述（由申请人提供）：该研究计划的总体主旨是开发新的和改进的方法来操纵和询问大分子离子，重点关注多肽种类，包括来自酶促或化学消化的肽、全蛋白质和蛋白质复合物。最终目标是最大化从感兴趣的蛋白质和蛋白质复合物中获得的结构信息，以用于其鉴定和表征。这项工作的重点是离子形成、离子化学以及与质谱和串联质谱相关的仪器。气相离子充当感兴趣的凝聚相物质的替代物。串联质谱中的结构信息来源于离子化学和带电反应产物质量的精确测量。离子的性质（例如，质子化分子与自由基阳离子）在其化学过程中起着重要作用，反应条件也是如此。这项研究的前提是新离子化学、新工具和新方法可以解决目前质谱从大型多肽物种中获取结构信息的能力的许多限制。它还基于这样的前提：将电离过程与离子操作/询问和质量分析步骤分开是可取的并且是可能的，以便每个步骤都可以单独优化。为了进一步增加通过质谱和串联质谱获得的蛋白质结构信息，我们提出了以下具体目标： 具体目标 1：用于蛋白质鉴定和结构表征的新的和改进的基于离子化学的方法。具体目标 2：在“混合”串联质谱仪中对肽和蛋白质进行串联质谱分析的新仪器和方法。具体目标 3：开发通过基质辅助激光解吸电离 (MALDI) 电离蛋白质的串联质谱新方法。这些目标将通过开发用于离子转化的新型离子/离子反应、扩大离子活化方法的范围、支持新离子化学的仪器开发以及开发通过离子化学和仪器的进步实现的蛋白质识别/表征新方法来实现。公共卫生相关性：该项目旨在改进蛋白质鉴定/表征。识别蛋白质并表征其翻译后修饰（即修饰的身份和位置）的能力是许多基础生物医学研究的基础。这种能力也是识别蛋白质生物标志物及其与疾病相关性的关键。该项目旨在通过极大地扩展通过质谱可访问的信息内容来解决这一任务。