Development of Methods for Rapid Proteomic Analysis Using Accurate Mass Measurement

利用精确质量测量开发快速蛋白质组分析方法

• 批准号: 0316002
• 负责人: I. Jonathan Amster
• 金额: --
• 依托单位: University of Georgia Research Foundation Inc
• 依托单位国家: 美国
• 项目类别: Continuing Grant
• 财政年份: 2003
• 资助国家: 美国
• 起止时间: 2003-07-15 至 2007-06-30
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=0316002&HistoricalAwards=false
• 关键词: Development; Methods; Rapid; Proteomic; Analysis

Professor Jonathan Amster of the University of Georgia is supported by the Analytical and Surface Chemistry Program to develop new methods for quantitatively analyzing changes in protein expression in biological systems. The goal is to be able to analyze all proteins in a mixture simultaneously, thus providing a substantial reduction in analysis time and effort. Mixtures of proteins will be enzymatically digested, separated by liquid chromatography and analyzed by high resolution mass spectrometry. Proteins in the original mixture will be identified from their proteolytic fragments by using the accurate mass data. "Mass defect labeling" is being invented as a way to increase the specificity of the assignment. The mass defect tags shift the peptide mass to a "clear" region of the m/z plot where they can be easily distinguished from nearby untagged peaks. Several novel reagents are proposed and are being synthesized as part of the project. Additional experiments will explore the use of endogenous labeling with a stable isotope in concert with the use of mass defects. The work represents a high throughput approach to the analysis of the proteome, meaning the library of proteins expressed by an organism. In addition, the work will impact other arenas of mass spectrometry including environmental and biomedical research.

乔治亚大学的Jonathan Amster教授在分析和表面化学项目的支持下，开发了定量分析生物系统中蛋白质表达变化的新方法。目标是能够同时分析混合物中的所有蛋白质，从而大大减少分析时间和工作量。蛋白质混合物将被酶消化，用液相色谱分离，并用高分辨率质谱分析。原始混合物中的蛋白质将通过使用精确的质量数据从其蛋白水解片段中识别出来。发明“质量缺陷标记”是为了增加分配的特异性。质量缺陷标签将肽质量转移到m/z图的“清晰”区域，在那里它们可以很容易地与附近未标记的峰区区分开来。作为该项目的一部分，提出并正在合成几种新型试剂。额外的实验将探索使用稳定同位素的内源性标记与使用质量缺陷相一致。这项工作代表了蛋白质组分析的高通量方法，蛋白质组是指生物体表达的蛋白质库。此外，这项工作将影响质谱法的其他领域，包括环境和生物医学研究。