Trailing end-point phenotype antibiotic-sensitive strains of Candida albicans produce different amounts of aspartyl peptidases.
Candida albicans is an opportunistic fungal pathogen that causes severe systemic infections in immunosuppressed individuals. C. albicans resistance to antifungal drugs is a severe problem in patients receiving prolonged therapy. Moreover, trailing yeast growth, which is defined as a resistant MIC after 48 h of incubation with triazole antifungal agents but a susceptible MIC after 24 h, has been noted in tests of antifungal susceptibility against some C. albicans isolates. In this context, we recently noticed this phenomenon in our routine susceptibility tests with fluconazole/itraconazole and C. albicans clinical isolates. In the present study, we investigated the production of cell-associated and secreted aspartyl peptidases (Saps) in six trailing clinical isolates of C. albicans, since this class of hydrolytic enzymes is a well-known virulence factor expressed by this fungal pathogen. Sap2, which is the best-studied member of the Sap family, was detected by flow cytometry on the cell surface of yeasts and as a 43-kDa polypeptide in the culture supernatant, as demonstrated by Western blotting assay using an anti-Sap1-3 polyclonal antibody. Released aspartyl peptidase activity was measured with BSA hydrolysis and inhibited by pepstatin A, showing distinct amounts of proteolytic activity ranging from 5.7 (strain 44B) to 133.2 (strain 11) arbitrary units. Taken together, our results showed that trailing clinical isolates of C. albicans produced different amounts of both cellular and secreted aspartyl-type peptidases, suggesting that this phenotypic feature did not generate a regular pattern regarding the expression of Sap.
Braga-Silva
L A
LA
Laboratório de Estudos Integrados em Bioquímica Microbiana, Departamento de Microbiologia Geral, Universidade Federal do Rio de Janeiro, Rio de Janeiro, RJ, Brasil.
Mesquita
D G A
DG
Ribeiro
M D
MD
Carvalho
S M F
SM
Fracalanzza
S E L
SE
Santos
A L S
AL
D000328:Adult; D000368:Aged; D000935:Antifungal Agents / Q000494:pharmacology*; D016282:Aspartic Acid Endopeptidases / Q000032:analysis* / Q000187:drug effects; D002176:Candida albicans / Q000187:drug effects / Q000201:enzymology* / Q000254:growth & development; D002675:Child, Preschool; D005260:Female; D015725:Fluconazole / Q000494:pharmacology*; D006801:Humans; D017964:Itraconazole / Q000494:pharmacology*; D008297:Male; D008826:Microbial Sensitivity Tests; D008875:Middle Aged; D010641:Phenotype; D012710:Serum Albumin, Bovine
D000935:Antifungal Agents; D012710:Serum Albumin, Bovine; D017964:Itraconazole; D015725:Fluconazole; D016282:Aspartic Acid Endopeptidases; C086390:SAP1 protein, Candida albicans