Differential Expression Proteomics: Identification and Quantitation of Peptides and Proteins by Fixed Charge Derivatization and Tandem Mass Spectrometry

这项提议的目的是开发新的策略，用于快速、灵敏和选择性地识别和定量复杂混合物中存在的蛋白质。具体地说，将开发含有选定氨基酸的多肽的同位素标记固定电荷衍生物，以引导串联质谱仪后产物离子的形成走向单一的裂解通道。这种方法将提供比现有方法更高的选择性和高达2个数量级的灵敏度，并将允许在蛋白质水平上检查细胞从正常状态转变为疾病状态后发生的复杂细胞变化。

The aim of this proposal is to develop novel strategies for the rapid, sensitive and selective identification and quantitation of proteins present in complex mixtures. Specifically, isotopically labeled fixed charge derivatives of peptides containing selected amino acids will be developed that direct the formation of product ions following tandem mass spectrometry toward a single fragmentation channel. This approach will provide enhanced selectivity and sensitivity of up to 2 orders of magnitude over existing approaches, and will allow examination, at the protein level, of the complex cellular changes that occur following transformation of cells from a normal to a diseased state.