Cited by 56Advanced Proteomics Techniques and ApplicationsMicrofluidic and Capillary Electrophoresis ApplicationsMass Spectrometry Techniques and Applications

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

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Zhao, Yimeng, Sun, Liangliang, Champion, Matthew M., Knierman, Michael D., Dovichi, Norman J.·2014·Analytical Chemistry

Abstract

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.

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  1. Kelleher N. L. Anal. Chem. 2004, 76, 197A– 203A. 15190879
  2. Kelleher N. L.; Lin H. Y.; Valaskovic G. A.; Aaserud D. J.; Fridriksson E. K.; McLafferty F. W. J. Am. Chem. Soc. 1999, 121, 806– 812.
  3. Qiao Y.; Di Z.; Ma Y.; Ma P.; Xia S. Chin. J. Chem. Eng. 2010, 18, 446– 454.

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Authors
Zhao, Y., Sun, L., Champion, M. M., Knierman, M. D., & Dovichi, N. J.
Affiliations
  • University of Notre Dame (US)
  • Eli Lilly (United States) (US)
  • University of Notre Dame , , Notre Dame , ,
Venue
Analytical Chemistry
Published
2014-04-11
Volume
86
Issue
10
Pages
4873-4878
ISSN
  • 0003-2700 (ISSN-L)
  • 1520-6882
OpenAlex
W2321501735
Cited by
56
References
36
Citations by year
2025: 3 · 2024: 2 · 2023: 2 · 2022: 2 · 2021: 5 · 2020: 2 · 2019: 4 · 2018: 5 · 2017: 6 · 2016: 10 · 2015: 11 · 2014: 4
FWCI
4.42
Type
journal article
Language
en
Open access
Locations
  • Analytical Chemistry (publishedVersion, other-oa)
  • PubMed Central (publishedVersion)
  • PubMed (publishedVersion)
Funders
National Institutes of Health (R01GM096767)
Topics
  • Physical Sciences › Chemistry › Spectroscopy › Advanced Proteomics Techniques and Applications
  • Physical Sciences › Engineering › Biomedical Engineering › Microfluidic and Capillary Electrophoresis Applications
  • Physical Sciences › Chemistry › Spectroscopy › Mass Spectrometry Techniques and Applications
Keywords
Mycobacterium marinum, secretome, top-down proteomics, capillary zone electrophoresis, proteoforms, post-translational modification, conductivity, culture filtrate, signal peptide removal, N-terminal methionine excision, acetylation
MeSH terms
Bacterial Proteins, Peptides, Protein Processing, Post-Translational, Electrophoresis, Capillary, Mycobacterium marinum, Spectrometry, Mass, Electrospray Ionization, Tandem Mass Spectrometry

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