Rapid identification of E. coli bacteriophages using mass spectrometry

Serafim, Vlad and Pantoja, Leonardo and Ring, Christopher J. and Shah, H. and Shah, Ajit J. (2017) Rapid identification of E. coli bacteriophages using mass spectrometry. Journal of Proteomics and Enzymology, 6 (1). pp. 1-5. ISSN 2470-1289

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Abstract

Objective: The current increasing interest in the application of mass spectrometry, in particular MALDI-TOF MS, to identification of bacteria and fungi calls for a need to utilise this technology for identification of other infectious agents such as viruses. The aim of the present study was to develop a rapid and reliable mass spectrometry-based proteomic method for identification of Escherichia coli phages.
Methods: The approach was based on rapid in-solution tryptic digestion of suspensions of plaque-purified bacteriophage followed by mass spectral analysis. Matrix-assisted laser desorption ionisation time-of-flight mass spectrometry (MALDI-TOF MS) and liquid chromatography – tandem mass spectrometry (LC-MS) were used to analyse the tryptic digests. Processing of tandem mass spectrometry data and interpretation of results were achieved using Mascot software and the Swiss-Prot database.
Results: Five bacteriophage species (Enterobacteria phages P2, T4, T5, T7 and Lambda) isolated in E. coli cultures were identified. The viral proteins were identified from a complex mixture of host bacterial proteins. In addition, using a single ion monitoring method, a Lambda prophage derived protein was also identified.
Conclusion: The data obtained demonstrate that LC-MS/MS can be used for accurate identification of E.coli- specific bacteriophages in both lytic and lysogenic cycles
Keywords: Bacteriophage virus; Mass-spectrometry; Liquid chromatography; MALDI; LC-MS/MS; Lytic; Lysogenic; Enterobacteria; E.coli; Phage; Viruses

Item Type: Article
Research Areas: A. > School of Science and Technology > Natural Sciences
Item ID: 22059
Notes on copyright: Article is published under open access
Useful Links:
Depositing User: Ajit Shah
Date Deposited: 16 Jun 2017 15:59
Last Modified: 05 Sep 2018 13:00
URI: http://eprints.mdx.ac.uk/id/eprint/22059

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