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学科主题: 物理化学
题名: 1-Dodecyl-3-Methylimidazolium Chloride-Assisted Sample Preparation Method for Efficient Integral Membrane Proteome Analysis
作者: Zhao, Qun1, 2;  Fang, Fei1, 2;  Liang, Yu1;  Yuan, Huiming1;  Yang, Kaiguang1;  Wu, Qi1, 2;  Liang, Zhen1;  Zhang, Lihua1;  Zhang, Yukui1
通讯作者: 张丽华
刊名: ANALYTICAL CHEMISTRY
发表日期: 2014-08-05
DOI: 10.1021/ac5013267
卷: 86, 期:15, 页:7544-7550
收录类别: SCI
文章类型: Article
WOS标题词: Science & Technology ;  Physical Sciences
类目[WOS]: Chemistry, Analytical
研究领域[WOS]: Chemistry
英文摘要: Due to their extremely hydrophobic nature, the analysis of integral membrane proteins (IMPs) is of great challenge. Although various additives have been applied to improve the solubility of IMPs, they still suffer from low solubilization efficiency, incompatibility with trypsin digestion, or interference with MS detection. Herein, the systematic study on the effect of ionic liquid structure on membrane protein solubilization and trypsin biocompatibility was performed, based on which 1-dodecyl-3-methylimidazolium chloride (C12Im-Cl) was selected for the sample preparation of IMPs. Compared with other commonly used additives, such as sodium dodecyl sulfate (SDS), Rapigest, and methanol, C12Im-Cl showed the best performance. In addition, with a strong cation exchange trap column, it could be easily removed after trypsin digestion, which not only was beneficial to avoid protein precipitation during digestion but also had no adverse effect on LC-MS-based separation and detection. Such a C12Im-Cl-assisted sample preparation method was further applied to the membrane proteome analysis of rat brain. Compared with the SDS-assisted method, 1.4 and 3.5 times improvement on the identified IMP and hydrophobic peptide number were achieved (251 vs 178, and 982 vs 279). All these results demonstrated that the C12Im-Cl-assisted sample preparation method is of great promise to promote the large-scale membrane proteome profiling.
关键词[WOS]: TANDEM MASS-SPECTROMETRY ;  IONIC LIQUIDS ;  TRYPTIC DIGESTION ;  INFRARED-SPECTROSCOPY ;  LC-MS/MS ;  PROTEINS ;  SOLUBILIZATION ;  METHANOL ;  IDENTIFICATION ;  CHROMATOGRAPHY
语种: 英语
WOS记录号: WOS:000340081100057
Citation statistics: 
内容类型: 期刊论文
URI标识: http://cas-ir.dicp.ac.cn/handle/321008/144459
Appears in Collections:中国科学院大连化学物理研究所_期刊论文

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作者单位: 1.Chinese Acad Sci, Dalian Inst Chem Phys, Natl Chromatog R&A Ctr, Key Lab Separat Sci Analyt Chem, Dalian 116023, Liaoning, Peoples R China
2.Univ Chinese Acad Sci, Beijing 100039, Peoples R China

Recommended Citation:
Zhao, Qun,Fang, Fei,Liang, Yu,et al. 1-Dodecyl-3-Methylimidazolium Chloride-Assisted Sample Preparation Method for Efficient Integral Membrane Proteome Analysis[J]. ANALYTICAL CHEMISTRY,2014,86(15):7544-7550.
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