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(Referência obtida automaticamente do Web of Science, por meio da informação sobre o financiamento pela FAPESP e o número do processo correspondente, incluída na publicação pelos autores.)

iochemical characterization and analysis of gene expression of an alpha-mannosidase secreted by Paracoccidioides brasiliensi

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Autor(es):
Correa, Priscila C. [1] ; Fernandes, Fabricio F. [2] ; Costa, V, Marcelo ; Landgraf, Taise N. [3] ; Panunto-Castelo, Ademilson [4]
Número total de Autores: 5
Afiliação do(s) autor(es):
[1] Univ Sao Paulo, Ribeirao Preto Med Sch, Grad Program Basic & Appl Immunol, BR-14049900 Ribeirao Preto, SP - Brazil
[2] Univ Paulista, BR-14024270 Ribeirao Preto, SP - Brazil
[3] Anhanguera Educ, BR-13614370 Leme, SP - Brazil
[4] Costa, Marcelo, V, Univ Sao Paulo, Fac Philosophy Sci & Letters Ribeirao Preto, Dept Biol, Av Bandeirantes 3900, BR-14040901 Ribeirao Preto, SP - Brazil
Número total de Afiliações: 4
Tipo de documento: Artigo Científico
Fonte: Medical Mycology; v. 60, n. 2 FEB 2 2022.
Citações Web of Science: 0
Resumo

Paracoccidioidomycosis (PCM) is a systemic mycosis caused by fungi of the Paracoccidioides genus, being endemic in Latin America and with the highest number of cases in Brazil. Paracoccidioides spp. release a wide range of molecules, such as enzymes, which may be important for PCM establishment. Here, we identified the 85- and 90-kDa proteins from the supernatants of P. brasiliensis cultures as being an alpha-mannosidase. Because the expected mass of this alpha-mannosidase is 124.2-kDa, we suggest that the proteins were cleavage products. Indeed, we found an alpha-mannosidase activity in the culture supernatants among the excreted/secreted antigens (ESAg). Moreover, we determined that the enzyme activity was optimal in buffer at pH 5.6, at the temperature of 45oC, and with a concentration of 3 mM of the substrate p-NP-alpha-D-Man. Remarkably, we showed that the gene expression of this alpha-mannosidase was higher in yeasts than hyphae in three P. brasiliensis isolates with different virulence degrees that were grown in Ham's F12 synthetic medium for 15 days. But in complex media YPD and Fava Netto, the significantly higher gene expression in yeasts than in hyphae was seen only for the virulent isolate Pb18, but not for intermediate virulence Pb339 and low virulence Pb265 isolates. These results about the high expression of the alpha-mannosidase gene in the pathogenic yeast form of P. brasiliensis open perspectives for studying this alpha-mannosidase concerning the virulence of P. brasiliensis isolates. Lay summary Paracoccidioides brasiliensis causes deep mycosis, paracoccidioidomycosis. We determined for the first time the biochemical properties of an alpha-mannosidase released by this fungus. We suggest that the enzyme gene expression in the fungus is associated with fungal morphology, stress, and virulence. (AU)

Processo FAPESP: 17/01390-8 - Avaliação de plasmídeo vacinal contendo o gene da proteína de estresse térmico de 60kDa de Paracoccidioides brasiliensis como terapia contra a paracoccidiodomicose experimental em camundongos
Beneficiário:Ademilson Panunto-Castelo
Modalidade de apoio: Auxílio à Pesquisa - Regular