Capillary Zone Electrophoresis-Electrospray Ionization-Tandem Mass Spectrometry for Top-Down Characterization of the Mycobacterium marinum Secretome

被引：44

作者：

Zhao, Yimeng ^{[1
]}

Sun, Liangliang ^{[1
]}

Champion, Matthew M. ^{[1
]}

Knierman, Michael D. ^{[2
]}

Dovichi, Norman J. ^{[1
]}

机构：

[1] Univ Notre Dame, Dept Chem & Biochem, Notre Dame, IN 46556 USA

[2] Eli Lilly & Co, Indianapolis, IN 46225 USA

来源：

ANALYTICAL CHEMISTRY | 2014年 / 86卷 / 10期

基金：

美国国家卫生研究院;

关键词：

INTACT PROTEINS; MEMBRANE-PROTEINS; BOTTOM-UP; PROTEOMICS; INTERFACE; KDA; RESOLUTION; TIME;

D O I：

10.1021/ac500092q

中图分类号：

O65 [分析化学];

学科分类号：

070302 ; 081704 ;

摘要：

Capillary zone electrophoresis (CZE) with an electrokinetically pumped sheath-flow nanospray interface was coupled with a high-resolution Q-Exactive mass spectrometer for the analysis of culture filtrates from Mycobacterium marinum. We confidently identified 22 gene products from the wildtype M. marinum secretome in a single CZE-tandem mass spectrometry (MS/MS) run. A total of 58 proteoforms were observed with post-translational modifications including signal peptide removal, N-terminal methionine excision, and acetylation. The conductivities of aqueous acetic acid and formic acid solutions were measured from 0.1% to 100% concentration (v/v). Acetic acid (70%) provided lower conductivity than 0.25% formic acid and was evaluated as low ionic-strength and a CZE-MS compatible sample buffer with good protein solubility.

引用

页码：4873 / 4878

页数：6

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