INSTRUMENTATION FOR ANALYSIS OF HIGH MASS BIOMOLECULES

描述（由申请人提供）：本研究计划的总体目标是开发新的和改进的方法，用于操纵和询问大分子离子，重点是多肽物质，包括酶促或化学消化产生的肽、完整蛋白质和蛋白质复合物。最终目标是最大限度地提高结构信息，可以从蛋白质和蛋白质复合物的目的，他们的鉴定和表征。工作重点涉及离子形成、离子化学和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自离子化学和带电反应产物质量的准确测量。离子的性质（例如，质子化分子对自由基阳离子）在其化学中起主要作用，反应条件也是如此。这项研究的前提是，新的离子化学，新的工具和新的方法可以解决许多目前的限制，质谱分析的能力，从大的多肽物种的结构信息。它也是基于这样的前提，即它是既可取的和可能的，从离子操作/询问和质量分析步骤，使每个可以单独优化的电离过程。为了进一步增加蛋白质的结构信息可从质谱和串联质谱，我们提出了以下具体目标：具体目标1：新的和改进的离子化学为基础的方法，蛋白质的鉴定和结构表征。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：基质辅助激光解吸电离（MALDI）蛋白质串联质谱新方法的开发。这些目标将通过开发用于离子转化的新型离子/离子反应、扩大范围的离子活化方法、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鉴定/表征的新方法来实现。公共卫生相关性：该项目旨在改进蛋白质鉴定/表征。鉴定蛋白质和表征其翻译后修饰的能力（即，修饰的身份和位置）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可通过质谱法获得的信息内容来解决这一任务。

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.