title: Proteomic profile of susceptible and multidrug-resistant clinical isolates of Escherichia coli and Klebsiella pneumoniae using label-free and immunoproteomic strategies.
abstract: Infectious diseases caused by multidrug-resistant (MDR) Enterobacteriaceae have exponentially increased in the past decade, and are a major concern in hospitals. In the first part of the work, we compared the proteome profile of MDR and susceptible clinical isolates of Escherichia coli and Klebsiella pneumoniae in order to identify possible biological processes associated with drug resistance and susceptible phenotypes, using a label-free approach. In the second part, we used an immunoproteomics approach to identify immunoreactive proteins in the same isolates. A total of 388 and 377 proteins were identified in MDR and susceptible E. coli, respectively, evidencing that biological processes related to translation are upregulated in E. coli MDR, while there is an upregulation of processes related to catalytic activity in K. pneumoniae MDR. Both MDR strains show downregulation of processes related to amino acid activation and tRNA amino-acylation. Our data also suggest that MDR strains have higher immunoreactivity than the susceptible strains. The application of high-throughput mass spectrometry (MS) and bioinformatics to the study of modulation of biological processes might shed light on the characterization of multidrug resistance in bacteria.
authors.affiliation: QOPNA, Mass Spectrometry Center, Department of Chemistry, University of Aveiro, Aveiro, Portugal; iBiMED - Institute for Biomedicine, University of Aveiro, Aveiro, Portugal.
authors.forename: Sandra
authors.initials: S
authors.lastname: Magalhães
authors.affiliation: iBiMED - Institute for Biomedicine, University of Aveiro, Aveiro, Portugal.
authors.forename: Miguel
authors.initials: M
authors.lastname: Aroso
authors.affiliation: Clinical Pathology, Centro Hospitalar do Baixo Vouga, Oliveira do Bairro, Portugal.
authors.forename: Inês
authors.initials: I
authors.lastname: Roxo
authors.affiliation: Clinical Pathology, Centro Hospitalar do Baixo Vouga, Oliveira do Bairro, Portugal; Institute of Education and Citizenship, Oliveira do Bairro, Portugal.
authors.forename: Sónia
authors.initials: S
authors.lastname: Ferreira
authors.affiliation: Clinical Pathology, Centro Hospitalar do Baixo Vouga, Oliveira do Bairro, Portugal.
authors.forename: Frederico
authors.initials: F
authors.lastname: Cerveira
authors.affiliation: Clinical Pathology, Centro Hospitalar do Baixo Vouga, Oliveira do Bairro, Portugal.
authors.forename: Elmano
authors.initials: E
authors.lastname: Ramalheira
authors.affiliation: QOPNA, Mass Spectrometry Center, Department of Chemistry, University of Aveiro, Aveiro, Portugal.
authors.forename: Rita
authors.initials: R
authors.lastname: Ferreira
authors.affiliation: iBiMED - Institute for Biomedicine, University of Aveiro, Aveiro, Portugal; Department of Physiology and Cardiothoracic Surgery, Faculty of Medicine, University of Porto, Porto, Portugal. Electronic address: rvitorino@ua.pt.
authors.forename: Rui
authors.initials: R
authors.lastname: Vitorino
mesh_terms: D000368:Aged; D000369:Aged, 80 and over; D049148:Aminoacylation; D001426:Bacterial Proteins / Q000276:immunology* / Q000302:isolation & purification* / Q000378:metabolism; D024901:Drug Resistance, Multiple, Bacterial / Q000235:genetics*; D004926:Escherichia coli / Q000187:drug effects / Q000235:genetics* / Q000276:immunology / Q000378:metabolism*; D004927:Escherichia coli Infections / Q000382:microbiology; D005260:Female; D006801:Humans; D007710:Klebsiella Infections / Q000382:microbiology; D007711:Klebsiella pneumoniae / Q000187:drug effects / Q000235:genetics* / Q000276:immunology; D008297:Male; D013058:Mass Spectrometry; D008826:Microbial Sensitivity Tests; D008875:Middle Aged; D040901:Proteomics / Q000379:methods*; D012343:RNA, Transfer; D015854:Up-Regulation; D001618:beta-Lactamases / Q000096:biosynthesis
chemical_list: D001426:Bacterial Proteins; D012343:RNA, Transfer; D001618:beta-Lactamases
keywords: Enterobacteriaceae; Immunoproteomics; Multidrug resistance; Quantitative proteomics