Sheathless Capillary Electrophoresis-Tandem Mass Spectrometry for Top-Down Characterization of Pyrococcus furiosus Proteins on a Proteome Scale

被引：61

作者：

Han, Xuemei ^{[1
]}

Wang, Yueju ^{[1
]}

Aslanian, Aaron ^{[1
,2
]}

Bern, Marshall ^{[3
,4
]}

Lavallee-Adam, Mathieu ^{[1
]}

Yates, John R., III ^{[1
]}

机构：

[1] Scripps Res Inst, Dept Chem Physiol, La Jolla, CA 92037 USA

[2] Salk Inst Biol Studies, Mol & Cell Biol Lab, La Jolla, CA 92037 USA

[3] Prot Metr, San Carlos, CA 94070 USA

[4] Xerox Corp, Palo Alto Res Ctr, Palo Alto, CA 94304 USA

来源：

ANALYTICAL CHEMISTRY | 2014年 / 86卷 / 22期

关键词：

ELECTRON-CAPTURE DISSOCIATION; ZONE-ELECTROPHORESIS; INTACT PROTEINS; ESCHERICHIA-COLI; PEPTIDE ANALYSIS; CE-MS; IDENTIFICATION; INTERFACE; KDA; CHROMATOGRAPHY;

D O I：

10.1021/ac503439n

中图分类号：

O65 [分析化学];

学科分类号：

070302 ; 081704 ;

摘要：

Intact protein analysis via top-down mass spectrometry (MS) provides the unique capability of fully characterizing protein isoforms and combinatorial post-translational modifications (PTMs) compared to the bottom-up MS approach. Front-end protein separation poses a challenge for analyzing complex mixtures of intact proteins on a proteomic scale. Here we applied capillary electrophoresis (CE) through a sheathless capillary electrophoresis-electrospray ionization (CESI) interface coupled to an Orbitrap Elite mass spectrometer to profile the proteome from Pyrococcus furiosus. CESI-top-down MS analysis of Pyrococcus furiosus cell lysate identified 134 proteins and 291 proteoforms with a total sample consumption of 270 ng in 120 min of total analysis time. Truncations and various PTMs were detected, including acetylation, disulfide bonds, oxidation, glycosylation, and hypusine. This is the largest scale analysis of intact proteins by CE-top-down MS to date.

引用

页码：11006 / 11012

页数：7

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