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Microb Pathog ; 192: 106706, 2024 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-38763316

RESUMO

Mannheimiahaemolytica is an opportunistic agent of the respiratory tract of bovines, a member of the Pasteurellaceae family, and the causal agent of fibrinous pleuropneumonia. This bacterium possesses different virulence factors, allowing it to colonize and infect its host. The present work describes the isolation and characterization of a serine protease secreted by M. haemolytica serotype 1. This protease was isolated from M. haemolytica cultured media by precipitation with 50 % methanol and ion exchange chromatography on DEAE-cellulose. It is a 70-kDa protease able to degrade sheep and bovine fibrinogen or porcine gelatin but not bovine IgG, hemoglobin, or casein. Mass spectrometric analysis indicates its identity with protease IV of M. haemolytica. The proteolytic activity was active between pH 5 and 9, with an optimal pH of 8. It was stable at 50 °C for 10 min but inactivated at 60 °C. The sera of bovines with chronic or acute pneumonia recognized this protease. Still, it showed no cross-reactivity with rabbit hyperimmune serum against the secreted metalloprotease from Actinobacilluspleuropneumoniae, another member of the Pasteurellaceae family. M. haemolytica secreted proteases could contribute to the pathogenesis of this bacterium through fibrinogen degradation, a characteristic of this fibrinous pleuropneumonia.


Assuntos
Fibrinogênio , Mannheimia haemolytica , Serina Proteases , Animais , Mannheimia haemolytica/enzimologia , Ovinos , Bovinos , Fibrinogênio/metabolismo , Concentração de Íons de Hidrogênio , Serina Proteases/metabolismo , Serina Proteases/isolamento & purificação , Temperatura , Proteólise , Peso Molecular , Gelatina/metabolismo , Estabilidade Enzimática , Proteínas de Bactérias/metabolismo , Proteínas de Bactérias/isolamento & purificação , Espectrometria de Massas , Cromatografia por Troca Iônica , Suínos , Fatores de Virulência/metabolismo , Fatores de Virulência/isolamento & purificação
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