title: Comparative proteomics of an extended spectrum β-lactamase producing Escherichia coli strain from the Iberian wolf.
abstract: The Iberian wolf (Canis lupus signatus) is an endangered species native to the Iberian Peninsula. Due to their predatory and wild nature, these wolves serve as important indicators of environmental contamination by antimicrobial-resistant bacteria. β-Lactam antibiotics like cefotaxime are the most commonly used antibacterial agents. Bacterial resistance to these antibiotics occurs predominantly through enzymatic inactivation by extended-spectrum beta-lactamases. Escherichia coli strain WA57, isolated from Iberian wolf feces, is a cefotaxime-resistant strain that produces extended-spectrum beta-lactamases. In this study, using 2D-GE combined with MS and bioinformatics, we report significant differences in the abundance of 40 protein spots (p<0.01) from the extracellular, periplasmic, cytoplasmic, and membrane sub-proteomes and the whole-cell proteome of WA57 exposed and non-exposed to cefotaxime. A total of 315 protein spots were collected for protein identification. The comparative proteomics presented gives an overview of the complex changes in expression and metabolism that occur when WA57 is stressed with cefotaxime. Abundance of chaperone, porin and export proteins is particularly affected showing that the stress response and transport functions might directly influence the antibiotic resistance of this strain.
  
  
  CONCLUSIONS
  This study highlights the importance of proteomics in detecting protein expression changes in bacterial strains exposed to stress such as that caused by cefotaxime. This approach might help us understand which pathways form barriers for antibiotics. This article is part of a Special Issue entitled: Environmental and structural proteomics.
authors.affiliation: Institute for Biotechnology and Bioengineering, Center of Genomics and Biotechnology, University of Trás-os-Montes and Alto Douro, Vila Real, Portugal; Department of Genetics and Biotechnology, University of Trás-os-Montes and Alto Douro, Vila Real, Portugal; Center for Animal Science and Veterinary, Vila Real, Portugal; Department of Veterinary Science, University of Trás-os-Montes and Alto Douro, Vila Real, Portugal.
authors.forename: A
authors.initials: A
authors.lastname: Gonçalves
authors.affiliation: Center for Animal Science and Veterinary, Vila Real, Portugal; Department of Veterinary Science, University of Trás-os-Montes and Alto Douro, Vila Real, Portugal.
authors.forename: P
authors.initials: P
authors.lastname: Poeta
authors.affiliation: Institute for Biotechnology and Bioengineering, Center of Genomics and Biotechnology, University of Trás-os-Montes and Alto Douro, Vila Real, Portugal; Department of Genetics and Biotechnology, University of Trás-os-Montes and Alto Douro, Vila Real, Portugal; Center for Animal Science and Veterinary, Vila Real, Portugal; Department of Veterinary Science, University of Trás-os-Montes and Alto Douro, Vila Real, Portugal.
authors.forename: R
authors.initials: R
authors.lastname: Monteiro
authors.affiliation: Institute for Biotechnology and Bioengineering, Center of Genomics and Biotechnology, University of Trás-os-Montes and Alto Douro, Vila Real, Portugal; Department of Genetics and Biotechnology, University of Trás-os-Montes and Alto Douro, Vila Real, Portugal; Center for Animal Science and Veterinary, Vila Real, Portugal; Department of Veterinary Science, University of Trás-os-Montes and Alto Douro, Vila Real, Portugal.
authors.forename: C
authors.initials: C
authors.lastname: Marinho
authors.affiliation: Center for Animal Science and Veterinary, Vila Real, Portugal.
authors.forename: N
authors.initials: N
authors.lastname: Silva
authors.affiliation: Department of Animal Biology, Centre for Environmental Biology, Faculty of Sciences, University of Lisbon, Lisboa, Portugal.
authors.forename: A
authors.initials: A
authors.lastname: Guerra
authors.affiliation: Department of Animal Biology, Centre for Environmental Biology, Faculty of Sciences, University of Lisbon, Lisboa, Portugal.
authors.forename: F
authors.initials: F
authors.lastname: Petrucci-Fonseca
authors.affiliation: Center for Animal Science and Veterinary, Vila Real, Portugal; Department of Veterinary Science, University of Trás-os-Montes and Alto Douro, Vila Real, Portugal.
authors.forename: J
authors.initials: J
authors.lastname: Rodrigues
authors.affiliation: Área de Bioquímica y Biología Molecular, Universidad de La Rioja, Logroño, Spain.
authors.forename: C
authors.initials: C
authors.lastname: Torres
authors.affiliation: Chemistry Department, University of Aveiro, Aveiro, Portugal.
authors.forename: R
authors.initials: R
authors.lastname: Vitorino
authors.affiliation: Chemistry Department, University of Aveiro, Aveiro, Portugal.
authors.forename: P
authors.initials: P
authors.lastname: Domingues
authors.affiliation: Institute for Biotechnology and Bioengineering, Center of Genomics and Biotechnology, University of Trás-os-Montes and Alto Douro, Vila Real, Portugal; Department of Genetics and Biotechnology, University of Trás-os-Montes and Alto Douro, Vila Real, Portugal. Electronic address: gigrejas@utad.pt.
authors.forename: G
authors.initials: G
authors.lastname: Igrejas
mesh_terms: D000818:Animals; D004926:Escherichia coli / Q000145:classification* / Q000302:isolation & purification / Q000378:metabolism*; D029968:Escherichia coli Proteins / Q000378:metabolism*; D055442:Metabolome / Q000502:physiology*; D020543:Proteome / Q000378:metabolism*; D013045:Species Specificity; D019596:Wolves / Q000145:classification / Q000382:microbiology*; D001618:beta-Lactamases / Q000378:metabolism*
chemical_list: D029968:Escherichia coli Proteins; D020543:Proteome; D001618:beta-Lactamases
keywords: Antibiotic resistance; Escherichia coli; Iberian wolf; Mass spectrometry; Subcellular proteomes