Coupling Formic Acid Assisted Solubilization and Online Immobilized Pepsin Digestion with Strong Cation Exchange and Microflow Reversed-Phase Liquid Chromatography with Electrospray Ionization Tandem Mass Spectrometry for Integral Membrane Proteome Analysis

被引：34

作者：

Ma, Junfeng ^{[1
,2
,3
]}

Hou, Chunyan ^{[1
,2
,3
]}

Sun, Liangliang ^{[1
,2
,3
]}

Tao, Dingyin ^{[1
,2
,3
]}

Zhang, Yanyan ^{[2
,3
]}

Shan, Yichu ^{[1
,2
]}

Liang, Zhen ^{[1
,2
]}

Zhang, Lihua ^{[1
,2
]}

Yang, Ling ^{[2
]}

Zhang, Yukui ^{[1
,2
]}

机构：

[1] Chinese Acad Sci, Dalian Inst Chem Phys, Key Lab Separat Sci Analyt Chem, Natl Chromatog Res & Anal Ctr, Dalian 116023, Peoples R China

[2] Chinese Acad Sci, Dalian Inst Chem Phys, Div Biotechnol, Dalian 116023, Peoples R China

[3] Chinese Acad Sci, Grad Sch, Beijing 100039, Peoples R China

来源：

ANALYTICAL CHEMISTRY | 2010年 / 82卷 / 23期

关键词：

PROTEINS; SURFACTANT; MS/MS; MS;

D O I：

10.1021/ac1023099

中图分类号：

O65 [分析化学];

学科分类号：

070302 ; 081704 ;

摘要：

In this study, a facile system for membrane proteome profiling was established, in which membrane proteins were solubilized by formic acid, online digested by a pepsin-based immobilized enzyme reactor (pepsin IMER), and analyzed by strong cation exchange and microflow reversed phase liquid chromatography with electrospray ionization tandem mass spectrometry (SCX-mu RPLC-ESI-MS/MS) Under optimized conditions, such a system showed excellent compatibility between all crucial steps and was successfully applied for analyzing integral membrane proteins extracted from rat liver microsomes Out of the 235 unique proteins positively identified, 39% (91/235) were annotated as membrane proteins with one or more transmembrane domains (TMDs) It is anticipated that the efficient sample treatment and the relevant online analytical system might provide a promising tool for automated and comprehensive profiling of membrane proteomes

引用

页码：9622 / 9625

页数：4

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