The expression of the secreted aspartyl proteinases Sap4 to Sap6 from Candida albicans in murine macrophages
Détails
ID Serval
serval:BIB_097CC08812BE
Type
Article: article d'un périodique ou d'un magazine.
Collection
Publications
Institution
Titre
The expression of the secreted aspartyl proteinases Sap4 to Sap6 from Candida albicans in murine macrophages
Périodique
Molecular Microbiology
ISSN
0950-382X (Print)
Statut éditorial
Publié
Date de publication
05/1998
Volume
28
Numéro
3
Pages
543-54
Notes
Journal Article
Research Support, Non-U.S. Gov't --- Old month value: May
Research Support, Non-U.S. Gov't --- Old month value: May
Résumé
Medically important yeasts of the genus Candida secrete aspartyl proteinases (Sap), which are of particular interest as virulence factors. Six closely related gene sequences, SAP1 to SAP6, for secreted proteinases are present in Candida albicans. The methylotrophic yeast Pichia pastoris was chosen as an expression system for preparing substantial amounts of each Sap isoenzyme. Interestingly, Sap4, Sap5 and Sap6, which have not yet been detected in C. albicans cultures in vitro, were produced as active recombinant enzymes. Different Sap polyclonal antibodies were raised in rabbits and tested before further application by enzyme-linked immunosorbent assay (ELISA) against each recombinant Sap. Two antisera recognized only Sap4 to Sap6. Using these antisera, together with sap null mutants obtained by targeted mutagenesis, we could demonstrate a high production of Sap4, Sap5 and Sap6 by C. albicans cells after phagocytosis by murine peritoneal macrophages. Furthermore, a delta sap4,5,6 null mutant was killed 53% more effectively after contact with macrophages than the wild-type strain. These results support a role for Sap4 to Sap6 in pathogenicity.
Mots-clé
Animals
Antibodies, Fungal/immunology
Antigens, Fungal/analysis
Aspartic Endopeptidases/genetics/immunology/isolation &
purification/*metabolism
Candida albicans/*enzymology/genetics/growth & development/pathogenicity
Cross Reactions
Enzyme-Linked Immunosorbent Assay
Fluorescent Antibody Technique
*Fungal Proteins
Hydrogen-Ion Concentration
Isoenzymes/metabolism
Macrophages, Peritoneal/*microbiology
Mice
Phagocytosis
Pichia/enzymology
Plasmids/genetics
Rabbits
Recombinant Fusion Proteins/isolation & purification/metabolism
Pubmed
Web of science
Open Access
Oui
Création de la notice
25/01/2008 17:47
Dernière modification de la notice
20/08/2019 13:31