Development of Methods for Rapid Proteomic Analysis Using Accurate Mass Measurement

格鲁吉亚大学的Jonathan Amster教授得到了分析和表面化学计划的支持，以开发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，从而大幅减少分析时间和工作量。 蛋白质混合物将被酶促消化，通过液相色谱法分离，并通过高分辨率质谱法分析。 通过使用准确的质量数据，从蛋白水解片段中鉴定原始混合物中的蛋白质。 “质量缺陷标记”是作为一种增加分配特异性的方法而发明的。 质量缺陷标签将肽质量转移到m/z图的“清晰”区域，在该区域中它们可以容易地与附近的未标记峰区分开。 作为该项目的一部分，提出并正在合成几种新试剂。 另外的实验将探索使用稳定同位素的内源性标记与质量缺陷的使用。 这项工作代表了蛋白质组分析的高通量方法，这意味着生物体表达的蛋白质库。 此外，这项工作还将影响质谱的其他领域，包括环境和生物医学研究。

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.