TY - JOUR
T1 - 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
AU - Ma, Junfeng
AU - Hou, Chunyan
AU - Sun, Liangliang
AU - Tao, Dingyin
AU - Zhang, Yanyan
AU - Shan, Yichu
AU - Liang, Zhen
AU - Zhang, Lihua
AU - Yang, Ling
AU - Zhang, Yukui
PY - 2010/12/1
Y1 - 2010/12/1
N2 - 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-μRPLC-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.
AB - 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-μRPLC-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.
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U2 - 10.1021/ac1023099
DO - 10.1021/ac1023099
M3 - Article
C2 - 21047061
AN - SCOPUS:78649691258
SN - 0003-2700
VL - 82
SP - 9622
EP - 9625
JO - Analytical Chemistry
JF - Analytical Chemistry
IS - 23
ER -