Moisés Martínez-Castillo1, Rosa Elena Cárdenas-Guerra1, Rossana Arroyo1, Anjan Debnath2, Mario Alberto Rodríguez1, Myrna Sabanero3, Fernando Flores-Sánchez4, Fernando Navarro-Garcia4, Jesús Serrano-Luna4, Mineko Shibayama1. 1. Department of Infectomics & Molecular Pathogenesis, Center for Research & Advanced Studies of the National Polytechnic Institute, Av IPN 2508, Mexico City 07360, Mexico. 2. Center for Discovery & Innovation in Parasitic Diseases, Skaggs School of Pharmacy & Pharmaceutical Sciences, University of California San Diego, La Jolla, CA 92093, USA. 3. Department of Biology, University of Guanajuato, Noria Alta S/N, Noria Alta, Guanajuato 36050, Mexico. 4. Department of Cell Biology, Center for Research & Advanced Studies of the National Polytechnic Institute, Av IPN 2508, Mexico City 07360, Mexico.
Abstract
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.
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.
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