Proteomic comparison of three clinical diarrhoeagenic drug-resistant Escherichia coli isolates grown on CHROMagar™STEC media

John Bosco Kalule, Suereta Fortuin, Bridget Calder, Lourens Robberts, Karen H. Keddy, Andrew J.M. Nel, Shaun Garnett, Mark Nicol, Digby F. Warner, Nelson C. Soares, Jonathan M. Blackburn

Research output: Contribution to journalArticle

5 Citations (Scopus)

Abstract

Shiga-toxin-producing Escherichia coli (STEC) and enteropathogenic Escherichia coli (EPEC) are key diarrhoea-causing foodborne pathogens. We used proteomics to characterize the virulence and antimicrobial resistance protein profiles of three clinical pathogenic E. coli isolates (two EPEC [one resistant to ciprofloxacin] and one STEC) cultured on CHROMagar™STEC solid media after minimal laboratory passage. We identified 4767 unique peptides from 1630 protein group across all three clinical E. coli strains. Label-free proteomic analysis allowed the identification of virulence and drug resistance proteins that were unique to each of the clinical isolates compared in this study. The B subunit of Shiga toxin, ToxB, was uniquely detected in the STEC strain while several other virulence factors including SheA, OmpF, OmpC and OmpX were significantly more abundant in the STEC strain. The ciprofloxacin resistant EPEC isolate possessed reduced levels of key virulence proteins compared to the ciprofloxacin susceptible EPEC and STEC strains. Parallel reaction monitoring assays validated the presence of biologically relevant proteins across biologically-replicated cultures. Propagation of clinical isolates on a relevant solid medium followed by mass spectrometry analysis represents a convenient means to quantify virulence factors and drug resistance determinants that might otherwise be lost through extensive in vitro passage in enteropathogenic bacteria. Significance: Through the use of quantitative proteomics, we have characterized the virulence and antimicrobial resistance attributes of three clinically isolated, pathogenic E. coli strains cultured on solid media. Our results provide new, quantitative data on the expressed proteomes of these tellurite-resistant, diarrhoeagenic E. coli strains and reveal a subset of antimicrobial resistance and virulence proteins that are differentially abundant between these clinical strains. Our quantitative proteomics-based approach should thus have applicability in microbiological diagnostic labs for the identification of pathogenic/drug resistant E. coli in the future.

Original languageEnglish
Pages (from-to)25-35
Number of pages11
JournalJournal of Proteomics
Volume180
DOIs
Publication statusPublished - 30 May 2018
Externally publishedYes

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Shiga-Toxigenic Escherichia coli
Proteomics
Escherichia coli
Enteropathogenic Escherichia coli
Virulence
Shiga Toxin
Ciprofloxacin
Pharmaceutical Preparations
Proteins
Virulence Factors
Drug Resistance
Proteome
Diarrhea
Mass Spectrometry
Bacteria
Peptides
Pathogens
Mass spectrometry
Labels
Assays

Cite this

Kalule, J. B., Fortuin, S., Calder, B., Robberts, L., Keddy, K. H., Nel, A. J. M., ... Blackburn, J. M. (2018). Proteomic comparison of three clinical diarrhoeagenic drug-resistant Escherichia coli isolates grown on CHROMagar™STEC media. Journal of Proteomics, 180, 25-35. https://doi.org/10.1016/j.jprot.2017.09.003
Kalule, John Bosco ; Fortuin, Suereta ; Calder, Bridget ; Robberts, Lourens ; Keddy, Karen H. ; Nel, Andrew J.M. ; Garnett, Shaun ; Nicol, Mark ; Warner, Digby F. ; Soares, Nelson C. ; Blackburn, Jonathan M. / Proteomic comparison of three clinical diarrhoeagenic drug-resistant Escherichia coli isolates grown on CHROMagar™STEC media. In: Journal of Proteomics. 2018 ; Vol. 180. pp. 25-35.
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Kalule, JB, Fortuin, S, Calder, B, Robberts, L, Keddy, KH, Nel, AJM, Garnett, S, Nicol, M, Warner, DF, Soares, NC & Blackburn, JM 2018, 'Proteomic comparison of three clinical diarrhoeagenic drug-resistant Escherichia coli isolates grown on CHROMagar™STEC media' Journal of Proteomics, vol. 180, pp. 25-35. https://doi.org/10.1016/j.jprot.2017.09.003

Proteomic comparison of three clinical diarrhoeagenic drug-resistant Escherichia coli isolates grown on CHROMagar™STEC media. / Kalule, John Bosco; Fortuin, Suereta; Calder, Bridget; Robberts, Lourens; Keddy, Karen H.; Nel, Andrew J.M.; Garnett, Shaun; Nicol, Mark; Warner, Digby F.; Soares, Nelson C.; Blackburn, Jonathan M.

In: Journal of Proteomics, Vol. 180, 30.05.2018, p. 25-35.

Research output: Contribution to journalArticle

TY - JOUR

T1 - Proteomic comparison of three clinical diarrhoeagenic drug-resistant Escherichia coli isolates grown on CHROMagar™STEC media

AU - Kalule, John Bosco

AU - Fortuin, Suereta

AU - Calder, Bridget

AU - Robberts, Lourens

AU - Keddy, Karen H.

AU - Nel, Andrew J.M.

AU - Garnett, Shaun

AU - Nicol, Mark

AU - Warner, Digby F.

AU - Soares, Nelson C.

AU - Blackburn, Jonathan M.

PY - 2018/5/30

Y1 - 2018/5/30

N2 - Shiga-toxin-producing Escherichia coli (STEC) and enteropathogenic Escherichia coli (EPEC) are key diarrhoea-causing foodborne pathogens. We used proteomics to characterize the virulence and antimicrobial resistance protein profiles of three clinical pathogenic E. coli isolates (two EPEC [one resistant to ciprofloxacin] and one STEC) cultured on CHROMagar™STEC solid media after minimal laboratory passage. We identified 4767 unique peptides from 1630 protein group across all three clinical E. coli strains. Label-free proteomic analysis allowed the identification of virulence and drug resistance proteins that were unique to each of the clinical isolates compared in this study. The B subunit of Shiga toxin, ToxB, was uniquely detected in the STEC strain while several other virulence factors including SheA, OmpF, OmpC and OmpX were significantly more abundant in the STEC strain. The ciprofloxacin resistant EPEC isolate possessed reduced levels of key virulence proteins compared to the ciprofloxacin susceptible EPEC and STEC strains. Parallel reaction monitoring assays validated the presence of biologically relevant proteins across biologically-replicated cultures. Propagation of clinical isolates on a relevant solid medium followed by mass spectrometry analysis represents a convenient means to quantify virulence factors and drug resistance determinants that might otherwise be lost through extensive in vitro passage in enteropathogenic bacteria. Significance: Through the use of quantitative proteomics, we have characterized the virulence and antimicrobial resistance attributes of three clinically isolated, pathogenic E. coli strains cultured on solid media. Our results provide new, quantitative data on the expressed proteomes of these tellurite-resistant, diarrhoeagenic E. coli strains and reveal a subset of antimicrobial resistance and virulence proteins that are differentially abundant between these clinical strains. Our quantitative proteomics-based approach should thus have applicability in microbiological diagnostic labs for the identification of pathogenic/drug resistant E. coli in the future.

AB - Shiga-toxin-producing Escherichia coli (STEC) and enteropathogenic Escherichia coli (EPEC) are key diarrhoea-causing foodborne pathogens. We used proteomics to characterize the virulence and antimicrobial resistance protein profiles of three clinical pathogenic E. coli isolates (two EPEC [one resistant to ciprofloxacin] and one STEC) cultured on CHROMagar™STEC solid media after minimal laboratory passage. We identified 4767 unique peptides from 1630 protein group across all three clinical E. coli strains. Label-free proteomic analysis allowed the identification of virulence and drug resistance proteins that were unique to each of the clinical isolates compared in this study. The B subunit of Shiga toxin, ToxB, was uniquely detected in the STEC strain while several other virulence factors including SheA, OmpF, OmpC and OmpX were significantly more abundant in the STEC strain. The ciprofloxacin resistant EPEC isolate possessed reduced levels of key virulence proteins compared to the ciprofloxacin susceptible EPEC and STEC strains. Parallel reaction monitoring assays validated the presence of biologically relevant proteins across biologically-replicated cultures. Propagation of clinical isolates on a relevant solid medium followed by mass spectrometry analysis represents a convenient means to quantify virulence factors and drug resistance determinants that might otherwise be lost through extensive in vitro passage in enteropathogenic bacteria. Significance: Through the use of quantitative proteomics, we have characterized the virulence and antimicrobial resistance attributes of three clinically isolated, pathogenic E. coli strains cultured on solid media. Our results provide new, quantitative data on the expressed proteomes of these tellurite-resistant, diarrhoeagenic E. coli strains and reveal a subset of antimicrobial resistance and virulence proteins that are differentially abundant between these clinical strains. Our quantitative proteomics-based approach should thus have applicability in microbiological diagnostic labs for the identification of pathogenic/drug resistant E. coli in the future.

KW - Antimicrobial resistance

KW - Electron microscopy

KW - Proteomics

KW - Shiga-toxin-producing E. coli

KW - Virulence

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U2 - 10.1016/j.jprot.2017.09.003

DO - 10.1016/j.jprot.2017.09.003

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