A Stable Isotopic Pre-digestion Labeling Method for Protein Quantitative Analysis Using Matrix-assisted Laser Desorption/ionization Mass Spectrometry

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	A Stable Isotopic Pre-digestion Labeling Method for Protein Quantitative Analysis Using Matrix-assisted Laser Desorption/ionization Mass Spectrometry
	Fang F(方芳) ; Zhang J(张菁) ; Zhang L(张立) ; Guo YL(郭寅龙)
刊名	Chin. J. Chem.
	2009
卷号	27 期号:12 页码:2397-2404
ISSN号	1001-604X
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
通讯作者	郭寅龙
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.
学科主题	分析化学
收录类别	SCI
原文出处	http://dx.doi.org/10.1002/cjoc.201090015
语种	英语
WOS记录号	WOS:000273627600016
公开日期	2013-02-25
内容类型	期刊论文
源URL	[http://202.127.28.38/handle/331003/19998]
专题	上海有机化学研究所_分析化学研究室
推荐引用方式 GB/T 7714	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.
APA	方芳,张菁,张立,&郭寅龙.(2009).A Stable Isotopic Pre-digestion Labeling Method for Protein Quantitative Analysis Using Matrix-assisted Laser Desorption/ionization Mass Spectrometry.Chin. J. Chem.,27(12),2397-2404.
MLA	方芳,et al."A Stable Isotopic Pre-digestion Labeling Method for Protein Quantitative Analysis Using Matrix-assisted Laser Desorption/ionization Mass Spectrometry".Chin. J. Chem. 27.12(2009):2397-2404.