title: Differential proteomic analysis of outer membrane enriched extracts of Bacteroides fragilis grown under bile salts stress.
abstract: Bacteroides fragilis is the most commonly isolated anaerobic bacteria from infectious processes. Several virulence traits contribute to the pathogenic nature of this bacterium, including the ability to tolerate the high concentrations of bile found in the gastrointestinal tract (GIT). The activity of bile salts is similar to detergents and may lead to membrane permeabilization and cell death. Modulation of outer membrane proteins (OMPs) is considered a crucial event to bile salts resistance. The primary objective of the current work was to identify B. fragilis proteins associated with the stress induced by high concentration of bile salts. The outer membrane of B. fragilis strain 638R was isolated after growth either in the presence of 2% conjugated bile salts or without bile salts. The membrane fractions were separated on SDS-PAGE and analyzed by ESI-Q/TOF tandem mass spectrometry. A total of 37 proteins were identified; among them nine were found to be expressed exclusively in the absence of bile salts whereas eight proteins were expressed only in the presence of bile salts. These proteins are related to cellular functions such as transport through membrane, nutrient uptake, and protein-protein interactions. This study demonstrates the alteration of OMPs composition in B. fragilis during bile salts stress resistance and adaptation to environmental changes. Proteomics of OMPs was also shown to be a useful approach in the identification of new targets for functional analyses.
authors.affiliation: Departamento de Microbiologia Médica, Instituto de Microbiologia Paulo de Góes/UFRJ, Laboratório de Biologia de Anaeróbios, Universidade Federal do Rio de Janeiro, Rio de Janeiro, RJ, Brazil.
authors.forename: Renata F
authors.initials: RF
authors.lastname: Boente
authors.affiliation: Departamento de Microbiologia Médica, Instituto de Microbiologia Paulo de Góes/UFRJ, Laboratório de Biologia de Anaeróbios, Universidade Federal do Rio de Janeiro, Rio de Janeiro, RJ, Brazil.
authors.forename: Heidi
authors.initials: H
authors.lastname: Pauer
authors.affiliation: Departamento de Microbiologia Médica, Instituto de Microbiologia Paulo de Góes/UFRJ, Laboratório de Biologia de Anaeróbios, Universidade Federal do Rio de Janeiro, Rio de Janeiro, RJ, Brazil.
authors.forename: Deborah N S
authors.initials: DN
authors.lastname: Silva
authors.affiliation: Departamento de Microbiologia Médica, Instituto de Microbiologia Paulo de Góes/UFRJ, Laboratório de Biologia de Anaeróbios, Universidade Federal do Rio de Janeiro, Rio de Janeiro, RJ, Brazil.
authors.forename: Joaquim Santos
authors.initials: JS
authors.lastname: Filho
authors.affiliation: Instituto de Bioquímica Médica, Unidade de Espectrometria de Massas e Proteômica, Universidade Federal do Rio de Janeiro e Rede Proteômica do, Rio de Janeiro, RJ, Brazil.
authors.forename: Vanessa
authors.initials: V
authors.lastname: Sandim
authors.affiliation: Centro de Referência Prof. Hélio Fraga, Escola Nacional de Saúde Pública Sergio Arouca, Fundação Oswaldo Cruz, Rio de Janeiro, Brazil.
authors.forename: Luis Caetano M
authors.initials: LC
authors.lastname: Antunes
authors.affiliation: Departamento de Microbiologia Médica, Instituto de Microbiologia Paulo de Góes/UFRJ, Laboratório de Biologia de Anaeróbios, Universidade Federal do Rio de Janeiro, Rio de Janeiro, RJ, Brazil.
authors.forename: Rosana Barreto Rocha
authors.initials: RB
authors.lastname: Ferreira
authors.affiliation: Instituto de Bioquímica Médica, Unidade de Espectrometria de Massas e Proteômica, Universidade Federal do Rio de Janeiro e Rede Proteômica do, Rio de Janeiro, RJ, Brazil.
authors.forename: Russolina B
authors.initials: RB
authors.lastname: Zingali
authors.affiliation: Departamento de Microbiologia Médica, Instituto de Microbiologia Paulo de Góes/UFRJ, Laboratório de Biologia de Anaeróbios, Universidade Federal do Rio de Janeiro, Rio de Janeiro, RJ, Brazil.
authors.forename: Regina M C P
authors.initials: RM
authors.lastname: Domingues
authors.affiliation: Departamento de Microbiologia Médica, Instituto de Microbiologia Paulo de Góes/UFRJ, Laboratório de Biologia de Anaeróbios, Universidade Federal do Rio de Janeiro, Rio de Janeiro, RJ, Brazil. Electronic address: lobol@micro.ufrj.br.
authors.forename: Leandro A
authors.initials: LA
authors.lastname: Lobo
mesh_terms: D000222:Adaptation, Physiological; D001425:Bacterial Outer Membrane Proteins / Q000235:genetics / Q000302:isolation & purification* / Q000378:metabolism; D001441:Bacteroides fragilis / Q000737:chemistry / Q000187:drug effects* / Q000254:growth & development; D001647:Bile Acids and Salts / Q000494:pharmacology*; D002352:Carrier Proteins / Q000235:genetics / Q000302:isolation & purification* / Q000378:metabolism; D002462:Cell Membrane / Q000737:chemistry / Q000187:drug effects*; D003470:Culture Media / Q000737:chemistry; D015870:Gene Expression; D063990:Gene Ontology; D058977:Molecular Sequence Annotation; D040901:Proteomics / Q000379:methods; D013312:Stress, Physiological / Q000235:genetics*
chemical_list: D001425:Bacterial Outer Membrane Proteins; D001647:Bile Acids and Salts; D002352:Carrier Proteins; D003470:Culture Media
keywords: Bacteroides fragilis; Bile salts; ESI-Q/TOF; Outer membrane proteins; Resistance