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学科主题: 分析化学
题名: A Stable Isotopic Pre-digestion Labeling Method for Protein Quantitative Analysis Using Matrix-assisted Laser Desorption/ionization Mass Spectrometry
其他题名: 酶解前稳定同位素标记法结合基质辅助激光解析离子化质谱进行蛋白定量分析
作者: Fang F(方芳) ; Zhang J(张菁) ; Zhang L(张立) ; Guo YL(郭寅龙)
通讯作者: 郭寅龙
刊名: Chin. J. Chem.
发表日期: 2009
卷: 27, 期:12, 页:2397-2404
收录类别: SCI
部门归属: 上海有机所
英文摘要: As an extension of our previous work, here a strategy was demonstrated for protein identification and quantification analyses utilizing a combination of stable isotope chemical labeling with subsequent denaturation, enzymatic digestion and matrix assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS). Using [d0]- and [d6]-4,6-dimethoxy-2-(methylsulfonyl)pyrimidine ([d0]-/[d6]-DMMSP), stable isotopic labels were incorporated before digestion. The comparative samples were combined before labeling after digestion, thus biases resulting from differences in sample digestion were avoided and the higher accuracy of quantification could be attained. The labeling was spatial-selective to particular residues of cysteine, lysine, and tyrosine before denaturation, which could lead to a better universality of the strategy for cysteine-free proteins. In addition, some lysine residues were blocked after labeling, the partly destroyed recognition sites could simplify the trypsin hydrolysates and hence facilitate the MS complexity. Together, our one-step labeling strategy combined several desirable properties such as spatial-selective labeling, reliability of quantitative results, simplification of analysis of complex systems and direct analysis with minimum sample handling. Our results demonstrate the usefulness of the method for analyzing lysozyme in egg white. The method was expected to provide a new powerful tool for comparative proteome research.
语种: 英语
相关网址: 查看原文
WOS记录号: WOS:000273627600016
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内容类型: 期刊论文
URI标识: http://ir.sioc.ac.cn/handle/331003/19998
Appears in Collections:分析化学研究室_期刊论文

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Recommended Citation:
Fang F,Zhang J,Zhang L,et al. A Stable Isotopic Pre-digestion Labeling Method for Protein Quantitative Analysis Using Matrix-assisted Laser Desorption/ionization Mass Spectrometry[J]. Chin. J. Chem.,2009,27(12):2397-2404.
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