Could transformation mechanisms of acetylase-harboring pMdT1 plasmid be evaluated through proteomic tools in Escherichia coli?

  1. Magalhaes, P. 33
  2. Pinto, L. 33
  3. Gonçalves, A. 33
  4. Araújo, J.E. 8
  5. Santos, H.M. 78
  6. Capelo, J.L. 78
  7. Saenz, Y. 1
  8. de Toro, M. 16
  9. Torres, C. 12
  10. Chambon, C. 4
  11. Habraud, M. 45
  12. Poeta, P. 38
  13. Igrejas, G. 338
  1. 1 Centro de Investigación Biomédica de La Rioja
    info

    Centro de Investigación Biomédica de La Rioja

    Logroño, España

    ROR https://ror.org/03vfjzd38

  2. 2 Universidad de La Rioja
    info

    Universidad de La Rioja

    Logroño, España

    ROR https://ror.org/0553yr311

  3. 3 Universidade de Trás-os-Montes e Alto Douro
    info

    Universidade de Trás-os-Montes e Alto Douro

    Vila Real, Portugal

    ROR https://ror.org/03qc8vh97

  4. 4 Institut National de la Recherche Agronomique, Centre Auvergne-Rhône-Alpes, Plate-Forme d'Exploration du Métabolisme composante protéomique (PFEMcp), France
  5. 5 Institut National de la Recherche Agronomique, Centre Auvergne-Rhône-Alpes, UR454 Microbiologie, France
  6. 6 Universidad de Cantabria
    info

    Universidad de Cantabria

    Santander, España

    ROR https://ror.org/046ffzj20

  7. 7 ProteoMass Scientific Society, Faculty of Sciences and Technology, Caparica, Portugal
  8. 8 Universidade Nova de Lisboa
    info

    Universidade Nova de Lisboa

    Lisboa, Portugal

    ROR https://ror.org/02xankh89

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Revista:
Journal of Proteomics

ISSN: 1874-3919

Año de publicación: 2016

Volumen: 145

Páginas: 103-111

Tipo: Artículo

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DOI: 10.1016/J.JPROT.2016.03.042 SCOPUS: 2-s2.0-84964329097 WoS: WOS:000383931000011 GOOGLE SCHOLAR

Otras publicaciones en: Journal of Proteomics

Resumen

Escherichia coli is a commensal microorganism of the gastrointestinal tract of animals and humans and it is an excellent model organism for the study of antibiotic resistance mechanisms. The resistance transmission and other characteristics of bacteria are based on different types of gene transfer occurring throughout the bacterial evolution. One of which is horizontal gene transfer that allows us to understand the ability of bacteria to acquire new genes. One dimensional and two dimensional electrophoresis (2-DE) techniques were performed in order to identify and characterize the proteome of two E. coli strains: Electromax DH10B, a transformation-ready strain; and TF-Se20, the Electromax DH10B that contains the aac(6′)-Ib-cr4-harboring pMdT1 plasmid. After 2-DE and subsequent analysis by matrix-assisted laser desorption/ionization-time of flight mass spectrometry (MALDI-TOF MS), it was possible to identify 76 distinct proteins on the TF-Se20 strain, whereas 71 had a known function. From Electromax DH10B strain, 72 different proteins were identified of which 71 were associated with a biological process. The protein of interest, aminoglycoside N-(6′)-acetyltransferase type 1, was identified by MALDI-TOF MS. The liquid chromatography-tandem mass spectrometry (LC-MS/MS) technique was performed to determine its sequence. Seventy six percent of the acetylase sequence was reconstructed only in the TF-Se20 strain, representing the single protein associated to antibiotic resistance. MALDI-TOF MS and LC-MS/MS approaches allowed us to determine the total proteome of both strains, as well as the acetylase sequence. Both of them enhance the ability to obtain more accurate information about the mechanisms of antimicrobial resistance. The pMdT1 plasmid brings a new perspective in understanding the metabolic processes that lead to antibiotic resistance. Biological significance This study highlights the importance of proteomics and bioinformatics in understanding mechanisms of gene transfer and antibiotic resistance. These two approaches allow to compare the protein expression in different samples, as well as different biological processes related to each protein. © 2016 Elsevier B.V.