Nf-GH, a glycosidase secreted by Naegleria fowleri, causes mucin degradation: an in vitro and in vivo study

Nf-GH 是一种由福氏耐格里阿米巴原虫分泌的糖苷酶,可导致粘蛋白降解:一项体外和体内研究

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作者:Moisés Martínez-Castillo, Rosa Elena Cárdenas-Guerra, Rossana Arroyo, Anjan Debnath, Mario Alberto Rodríguez, Myrna Sabanero, Fernando Flores-Sánchez, Fernando Navarro-Garcia, Jesús Serrano-Luna, Mineko Shibayama

Aim

The aim of this work was to identify, characterize and evaluate the pathogenic role of mucinolytic activity released by Naegleria fowleri. Materials &

Conclusion

Our findings suggest that secretory products from N. fowleri play an important role in mucus degradation during the invasion process.

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.

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