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

Junfeng Ma, Chunyan Hou, Liangliang Sun, Dingyin Tao, Yanyan Zhang, Yichu Shan, Zhen Liang, Lihua Zhang, Ling Yang, Yukui Zhang

Research output: Contribution to journalArticle

Abstract

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.

Original languageEnglish (US)
Pages (from-to)9622-9625
Number of pages4
JournalAnalytical chemistry
Volume82
Issue number23
DOIs
StatePublished - Dec 1 2010

ASJC Scopus subject areas

  • Analytical Chemistry

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