title: Proteomic analysis of multidrug resistant Escherichia coli strains from scouring calves.
abstract: A number of researchers have used chemical inhibitors that target membrane efflux pumps as an experimental treatment strategy for multidrug resistant (MDR) bacterial infections. However, most of these compounds are toxic in vertebrate animals. The present research was therefore done to describe expression dynamics of drug resistance-associated Escherichia coli proteins that could serve as novel drug targets. Proteomes of MDR and antimicrobial susceptible (AS) E. coli were studied in two dimensional (2-D) polyacrylamide gels and liquid chromatography-mass spectrometry (LC-MS) was performed on proteins of interest. The number of recovered peptides per protein was used to elucidate the amounts of target proteins expressed in MDR and AS E. coli strains. Eight proteins that may be potentially involved in mechanisms of drug resistance were analyzed and identified by LC-MS. These were grouped into membrane porins (TolC, OmpA, OmpC, Nmpc Precursor), proteins involved in microbial protein synthesis (EF-Ts, EF-Tu, RpsA) and Dps, a protein of unknown location and function. Experimental data demonstrated variability in the expression patterns and quantities of the four porins (TolC, OmpA, OmpC, Nmpc precursor), the three microbial protein synthesis associated proteins (EF-Ts, EF-Tu and RpsA), and Dps which has been previously associated with drug resistance. While variability was seen in quantities and expression patterns of some of the proteins of interest, the present data falls short of determining the suitability of these proteins as novel drug targets. Further studies are required to explore how these proteins could be targeted for drug development.
authors.affiliation: Department of Veterinary and Microbiological Sciences, North Dakota State University, 1523 Centennial Blvd, Fargo, ND 58108, USA.
authors.forename: Ablesh
authors.initials: A
authors.lastname: Gautam
authors.forename: Heather M
authors.initials: HM
authors.lastname: Vinson
authors.forename: Penelope S
authors.initials: PS
authors.lastname: Gibbs
authors.forename: Susan
authors.initials: S
authors.lastname: Olet
authors.forename: Robert
authors.initials: R
authors.lastname: Barigye
mesh_terms: D000818:Animals; D002417:Cattle / Q000382:microbiology*; D002418:Cattle Diseases / Q000382:microbiology*; D002853:Chromatography, Liquid; D024901:Drug Resistance, Multiple, Bacterial; D015180:Electrophoresis, Gel, Two-Dimensional; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism*; D004927:Escherichia coli Infections / Q000382:microbiology / Q000662:veterinary*; D029968:Escherichia coli Proteins / Q000235:genetics / Q000378:metabolism; D013058:Mass Spectrometry; D018272:Porins / Q000235:genetics / Q000378:metabolism; D020543:Proteome / Q000032:analysis*; D040901:Proteomics / Q000379:methods
chemical_list: D029968:Escherichia coli Proteins; D018272:Porins; D020543:Proteome