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

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• 作者：Yimeng Zhao ; Liangliang Sun ; Matthew M. Champion ; Michael D. Knierman ; Norman J. Dovichi
• 刊名：Analytical Chemistry
• 出版年：2014
• 出版时间：May 20, 2014
• 年：2014
• 卷：86
• 期：10
• 页码：4873-4878
• 全文大小：303K
• 年卷期：v.86,no.10(May 20, 2014)
• ISSN：1520-6882

文摘

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鈥搕andem 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鈥揗S compatible sample buffer with good protein solubility.