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Yan X, Essaka DC, Sun L, et al. Bottom-up proteome analysis of E. coli using capillary zone electrophoresis-tandem mass spectrometry with an electrokinetic sheath-flow electrospray interface. Proteomics. 2013;13(17):2546-51doi: 10.1002/pmic.201300062.
Yan, X., Essaka, D. C., Sun, L., Zhu, G., & Dovichi, N. J. (2013). Bottom-up proteome analysis of E. coli using capillary zone electrophoresis-tandem mass spectrometry with an electrokinetic sheath-flow electrospray interface. Proteomics, 13(17), 2546-51. https://doi.org/10.1002/pmic.201300062
Yan, Xiaojing, et al. "Bottom-up proteome analysis of E. coli using capillary zone electrophoresis-tandem mass spectrometry with an electrokinetic sheath-flow electrospray interface." Proteomics vol. 13,17 (2013): 2546-51. doi: https://doi.org/10.1002/pmic.201300062
Yan X, Essaka DC, Sun L, Zhu G, Dovichi NJ. Bottom-up proteome analysis of E. coli using capillary zone electrophoresis-tandem mass spectrometry with an electrokinetic sheath-flow electrospray interface. Proteomics. 2013 Sep;13(17):2546-51. doi: 10.1002/pmic.201300062. Epub 2013 Aug 01. PMID: 23798545; PMCID: PMC4121376.
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Proteomics. 2013 Sep;13(17):2546-51. doi: 10.1002/pmic.201300062. Epub 2013 Aug 01.

Bottom-up proteome analysis of E. coli using capillary zone electrophoresis-tandem mass spectrometry with an electrokinetic sheath-flow electrospray interface.

Proteomics

Xiaojing Yan, David C Essaka, Liangliang Sun, Guijie Zhu, Norman J Dovichi

Affiliations

  1. Department of Chemistry and Biochemistry, University of Notre Dame, Notre Dame, IN, USA.

PMID: 23798545 PMCID: PMC4121376 DOI: 10.1002/pmic.201300062

Abstract

The Escherichia coli proteome was digested with trypsin and fractionated using SPE on a C18 SPE column. Seven fractions were collected and analyzed by CZE-ESI-MS/MS. The separation was performed in a 60-cm-long linear polyacrylamide-coated capillary with a 0.1% v/v formic acid separation buffer. An electrokinetic sheath-flow electrospray interface was used to couple the separation capillary with an Orbitrap-Velos operating in higher-energy collisional dissociation mode. Each CZE-ESI-MS/MS run lasted 50 min and total MS time was 350 min. A total of 23 706 peptide spectra matches, 4902 peptide IDs, and 871 protein group IDs were generated using MASCOT with false discovery rate less than 1% on the peptide level. The total mass spectrometer analysis time was less than 6 h, the sample identification rate (145 proteins/h) was more than two times higher than previous studies of the E. coli proteome, and the amount of sample consumed (<1 μg) was roughly fourfold less than previous studies. These results demonstrate that CZE is a useful tool for the bottom-up analysis of prokaryote proteomes.

© 2013 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

Keywords: Bottom-up proteomics; CZE-ESI-MS/MS; Electrokinetically driven sheath flow CE-MS interface; Escherichia coli; Technology

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