title: Expression of PZ-peptidases by cultures of several pathogenic fungi. Purification and characterization of a collagenase from Trichophyton schoenleinii.
abstract: Peptidolytic activity was studied in the broken-cell extracts of 17 isolates of pathogenic fungi tested with phenylazobenzyloxycarbonyl-Pro-Leu-Gly-Pro-Arg (PZ-PLGPA) as a substrate. All the fungi studied except Candida spp., Cryptococcus neoformans and two actinomycetes hydrolyzed the substrate and therefore contained a so-called PZ-peptidase activity. Of all the positive strains, Trichophyton schoenleinii, a pathogenic fungus showed the highest activity and was therefore chosen as a source for PZ-peptidase purification. The four chromatographic steps, a 'negative' dye column, a 'positive' dye column, hydroxyapatite Ultrogel, and modified TSK (HW 55), gave a highly purified peptidase with a 12% overall yield. Inhibitor studies suggested that the 82 000 M(r) PZ-peptidase is a metalloproteinase. Moreover it cleaved native rat type I collagen. Partial peptide sequencing showed a strong sequence homology to regions of two metalloproteinases previously identified in the yeast Saccharomyces cerevisiae and in rat.
authors.affiliation: Institut Pasteur, Unité de Mycologie, Paris, France.
authors.forename: O
authors.initials: O
authors.lastname: Ibrahim-Granet
authors.forename: F H
authors.initials: FH
authors.lastname: Hernandez
authors.forename: G
authors.initials: G
authors.lastname: Chevrier
authors.forename: B
authors.initials: B
authors.lastname: Dupont
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D003883:Arthrodermataceae / Q000201:enzymology; D017364:Collagenases / Q000302:isolation & purification; D005658:Fungi / Q000201:enzymology*; D008666:Metalloendopeptidases / Q000737:chemistry / Q000302:isolation & purification*; D008969:Molecular Sequence Data; D011480:Protease Inhibitors / Q000494:pharmacology; D051381:Rats; D017386:Sequence Homology, Amino Acid; D014249:Trichophyton / Q000201:enzymology; D015003:Yeasts
chemical_list: D011480:Protease Inhibitors; D017364:Collagenases; D008666:Metalloendopeptidases; C022510:thimet oligopeptidase