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JAC Advance Access originally published online on October 24, 2006
Journal of Antimicrobial Chemotherapy 2006 58(6):1145-1153; doi:10.1093/jac/dkl413
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© The Author 2006. Published by Oxford University Press on behalf of the British Society for Antimicrobial Chemotherapy. All rights reserved. For Permissions, please e-mail: journals.permissions@oxfordjournals.org

Effect of fluoroquinolone exposure on the proteome of Salmonella enterica serovar Typhimurium

Nick G. Coldham1,*, Luke P. Randall1, Laura J. V. Piddock2 and Martin J. Woodward1

1 Department of Food and Environmental Safety, Veterinary Laboratories Agency Addlestone, Surrey, KT15 3NB, UK 2 Antimicrobial Agents Research Group, Division of Immunity and Infection University of Birmingham, Birmingham, B15 2TT, UK

Received 6 July 2006; returned 8 August 2006; revised 5 September 2006; accepted 14 September 2006

*Corresponding author. Tel: +44-1932-357827; Fax: +44-1932-357595; E-mail: n.g.coldham{at}vla.defra.gsi.gov.uk

Objectives: The physiological response of Salmonella enterica serovar Typhimurium to fluoroquinolone antibiotics was investigated using proteomic methods.

Methods: Proteomes were prepared from strain SL1344 following treatment of broth cultures with ciprofloxacin (0.03 and 0.008 mg/L; 2x and 0.5x MIC) and enrofloxacin (0.03 mg/L) and from a multiple antibiotic resistant (MAR) mutant. Protein expression was determined by two-dimensional HPLC-MSn and also after exposure to ciprofloxacin by two-dimensional gel electrophoresis (2D-GE).

Results: The number of proteins (mean ± SD) detected by 2D-GE derived from control cultures of the wild-type strain was significantly (P < 0.05) reduced from 296 ± 77 to 153 ± 36 following treatment with ciprofloxacin (0.03 mg/L). Raised expression (P < 0.05) of 17 proteins was also detected, and increases of up to 8-fold (P < 0.0001) were observed for subunits of F1F0-ATP synthase, TolC and Imp. Analysis by two-dimensional HPLC-MSn provided higher proteome coverage with 787 ± 50 proteins detected, which was reduced (P < 0.005) to 560 ± 14 by ciprofloxacin (0.03 mg/L). Increased expression of 43 proteins was observed which included those detected by 2D-GE and additionally the efflux pump protein AcrB. The basal expression of the AcrAB/TolC efflux pump was elevated in the MAR mutant compared with the untreated wild-type and augmented following treatment with ciprofloxacin (0.03 mg/L). F1F0-ATP synthase and Imp were only elevated in the mutant when treated with ciprofloxacin.

Conclusions: These studies suggest that increased expression of AcrAB/TolC was associated with resistance while other increases, such as in F1F0-ATP synthase and Imp, were a response to fluoroquinolone.

Keywords: proteomics , fluoroquinolones , S. enterica


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