Cytosolic Neutral Proteinases of Paracoccidioides brasiliensis |
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Authors: | Gioconda San-Blas Françoise Sorais Gustavo Niño-Vega Cioly Méndez Felipe San-Blas |
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Institution: | (1) Instituto Venezolano de Investigaciones Cientificas (IVIC), Centro de Microbiología y Biología Celular, Apartado 21827, Caracas 1020A, Venezuela , VE |
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Abstract: | Cytosolic proteinases were assayed in both morphological phases of Paracoccidioides brasiliensis. Preparations from the mycelial phase were more active in vitro than those from the yeast cells. Optimal proteinase activities
for both phases occurred at pH's between 6.0 and 9.0, and at 45°C. Gelatin-SDS-PAGE electrophoresis separated several bands
(58–112 kDa) in mycelial preparations; a single band (70 kDa) was seen in yeast preparations. Enzymatic activities were inhibited
by antipain, phenyl methyl sulfonyl fluoride (PMSF), and chymostatin, suggestive of serine proteinases. Partial inhibition
of the mycelial enzymes by ethylene diamine tetraacetic acid (EDTA), 1,10-phenanthroline, and iodoacetamide, also suggested
the presence of cysteine- and metallo-proteinases. The enzymatic activity increased in preparations extracted from yeast cells
transforming to mycelia, and decreased in preparations obtained from the reverse process.
Received: 29 September 1997 / Accepted: 19 February 1998 |
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