RESUMO

AIM: The aim of this work was to identify, characterize and evaluate the pathogenic role of mucinolytic activity released by Naegleria fowleri. MATERIALS & METHODS: Zymograms, protease inhibitors, anion exchange chromatography, MALDI-TOF-MS, enzymatic assays, Western blot, and confocal microscopy were used to identify and characterize a secreted mucinase; inhibition assays using antibodies, dot-blots and mouse survival tests were used to evaluate the mucinase as a virulence factor. RESULTS: A 94-kDa protein with mucinolytic activity was inducible and abolished by p-hydroxymercuribenzoate. MALDI-TOF-MS identified a glycoside hydrolase. Specific antibodies against N. fowleri-glycoside hydrolase inhibit cellular damage and MUC5AC degradation, and delay mouse mortality. CONCLUSION: Our findings suggest that secretory products from N. fowleri play an important role in mucus degradation during the invasion process.

Assuntos

Glicosídeo Hidrolases/metabolismo , Mucinas/metabolismo , Naegleria fowleri/enzimologia , Fatores de Virulência/metabolismo , Animais , Western Blotting , Glicosídeo Hidrolases/química , Glicosídeo Hidrolases/efeitos dos fármacos , Humanos , Hidroximercuribenzoatos/farmacologia , Camundongos , Microscopia Confocal , Naegleria fowleri/efeitos dos fármacos , Naegleria fowleri/metabolismo , Naegleria fowleri/patogenicidade , Polissacarídeo-Liases/metabolismo , Espectrometria de Massas por Ionização e Dessorção a Laser Assistida por Matriz