title: Brucella abortus choloylglycine hydrolase affects cell envelope composition and host cell internalization.
abstract: Choloylglycine hydrolase (CGH, E.C. 3.5.1.24) is a conjugated bile salt hydrolase that catalyses the hydrolysis of the amide bond in conjugated bile acids. Bile salt hydrolases are expressed by gastrointestinal bacteria, and they presumably decrease the toxicity of host's conjugated bile salts. Brucella species are the causative agents of brucellosis, a disease affecting livestock and humans. CGH confers Brucella the ability to deconjugate and resist the antimicrobial action of bile salts, contributing to the establishment of a successful infection through the oral route in mice. Additionally, cgh-deletion mutant was also attenuated in intraperitoneally inoculated mice, which suggests that CGH may play a role during systemic infection other than hydrolyzing conjugated bile acids. To understand the role CGH plays in B. abortus virulence, we infected phagocytic and epithelial cells with a cgh-deletion mutant (Δcgh) and found that it is defective in the internalization process. This defect along with the increased resistance of Δcgh to the antimicrobial action of polymyxin B, prompted an analysis of the cell envelope of this mutant. Two-dimensional electrophoretic profiles of Δcgh cell envelope-associated proteins showed an altered expression of Omp2b and different members of the Omp25/31 family. These results were confirmed by Western blot analysis with monoclonal antibodies. Altogether, the results indicate that Brucella CGH not only participates in deconjugation of bile salts but also affects overall membrane composition and host cell internalization.
authors.affiliation: Instituto de Investigaciones Biotecnológicas-Instituto Tecnológico de Chascomús, Universidad Nacional de San Martín, Consejo Nacional de Investigaciones Científicas y Técnicas, Buenos Aires, Argentina.
authors.forename: María Inés
authors.initials: MI
authors.lastname: Marchesini
authors.forename: Joseph
authors.initials: J
authors.lastname: Connolly
authors.forename: María Victoria
authors.initials: MV
authors.lastname: Delpino
authors.forename: Pablo C
authors.initials: PC
authors.lastname: Baldi
authors.forename: Cesar V
authors.initials: CV
authors.lastname: Mujer
authors.forename: Vito G
authors.initials: VG
authors.lastname: DelVecchio
authors.forename: Diego J
authors.initials: DJ
authors.lastname: Comerci
mesh_terms: D000581:Amidohydrolases / Q000378:metabolism*; D000818:Animals; D001422:Bacterial Adhesion; D001425:Bacterial Outer Membrane Proteins / Q000378:metabolism; D002003:Brucella abortus / Q000201:enzymology* / Q000254:growth & development; D002006:Brucellosis / Q000382:microbiology; D002462:Cell Membrane / Q000378:metabolism*; D015169:Colony Count, Microbial; D004705:Endocytosis; D017353:Gene Deletion; D006367:HeLa Cells; D006801:Humans; D051379:Mice
chemical_list: D001425:Bacterial Outer Membrane Proteins; D000581:Amidohydrolases; C023154:choloylglycine hydrolase