Identification of major glucan-associated cell wall proteins of Candida albicans and their role in fluconazole resistance.
Identification of major glucan-associated proteins (GAPs) of the cell wall of a number of Candida albicans isolates susceptible or resistant to fluconazole (FLC) was addressed by direct sequencing of the protein bands resolved by unidimensional gel electrophoresis. Changes in the GAP compositions of the different strains grown in the presence of the drug were also investigated. In the FLC-susceptible strains, the major (more abundant) GAPs were enolase (46 kDa), two isoforms of phosphoglyceromutase (32 and 29 kDa), and two beta-(1-3)-exoglucanases (44 and 34 kDa), one of which (the 34-kDa component) was glycosylated. When these strains were grown in the presence of FLC there were substantial decreases in the intensities of the two enzymes of the glycolytic pathway (enolase and the phosphoglyceromutases), which were apparently replaced by enhancement of the exoglucanase constituents, particularly the 44-kDa one. This GAP pattern closely mimicked that observed in the FLC-resistant strains whether they were grown in the presence or in the absence of the drug. Both the enolase and the exoglucanase constituents were detected in the culture supernatants of FLC-treated cells, together with substantial amounts of highly glycosylated, probably mannoprotein secretory material, suggesting that FLC may cause marked alterations of GAP incorporation into the cell wall. Altogether, we were able to identify all major GAP constituents and monitor their distributions in the cell wall of C. albicans during treatment with FLC. The near equivalence of the GAP profile for the FLC-susceptible strain grown in the presence of FLC to that for the FLC-resistant strain suggests that the effects of the drug on GAPs may be stably incorporated into the cell wall of the fungus upon acquisition of resistance.
Angiolella
Letizia
L
Institute of Microbiology, Faculty of Pharmacy, Rome, Italy.
Micocci
Mia M
MM
D'Alessio
Simona
S
Girolamo
Antonietta
A
Maras
Bruno
B
Cassone
Antonio
A
D000935:Antifungal Agents / Q000494:pharmacology*; D015153:Blotting, Western; D002176:Candida albicans / Q000737:chemistry* / Q000187:drug effects*; D002241:Carbohydrates / Q000737:chemistry; D002473:Cell Wall / Q000737:chemistry*; D004352:Drug Resistance, Microbial; D004591:Electrophoresis, Polyacrylamide Gel; D015725:Fluconazole / Q000494:pharmacology*; D005656:Fungal Proteins / Q000737:chemistry*; D005936:Glucans / Q000737:chemistry*; D010751:Phosphopyruvate Hydratase / Q000037:antagonists & inhibitors / Q000276:immunology
D000935:Antifungal Agents; D002241:Carbohydrates; D005656:Fungal Proteins; D005936:Glucans; D015725:Fluconazole; D010751:Phosphopyruvate Hydratase