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Infectio
Print version ISSN 0123-9392
Abstract
GOMEZ, Salvador; GARCIA, Sandra Milena; DE BEDOUT, Catalina and GARCIA, Ana María. Protein profile analysis of Candida guilliermondii clinical isolates sensitive and resistant to fluconazole. Infect. [online]. 2011, vol.15, n.1, pp.20-24. ISSN 0123-9392.
Objectives: The aim of this study was to identify possible changes in the protein profiles of two isolates of Candida guilliermondii, one sensitive and other resistant to fluconazole to recognize proteins differentially expressed in relation to the resistance to this antimycotic. Methods: For this purpose, fractions from cytoplasm and membrane bound protein extracts from one resistant (MIC>256) and another sensitive (MIC=4) isolates were obtained and analyzed by SDS-PAGE. Results: Four protein bands present in the resistant isolation and absent in the sensitive isolate were identified. In the cytoplasmic extract three proteins were identified, one of 16kDa with the same size to YNK1p (Nucleoside diphosphate kinase), other of 37kDa with similar size to HEM13p (Coproporphyrinogen III oxidase) and/or ADHp1 (Alcohol Dehydrogenase) and the last of 45kDa was not identified previously in other proteomic studies made with other Candida species. In the membrane extract, one band corresponding to 25kDa protein HSP31p (Cysteine protease) was found. Conclusion: This is the first protein analysis study made in C. guilliermondii in which proteins potentially associated with resistance to fluconazole were found. In this research, four proteins bands differentially expressed and possibly associated to fluconazol resistance were identified. Three of these proteins, were previously described in other Candida species as related to azole resistance. The 45 kDa, hasnât been previously reported in other proteomic studies and could be specific to this species. Despite these results, further studies should be conducted on the protein profile of C. guilliermondii using more advanced and specific proteomic techniques as 2D-DIGE and mass spectrometry.
Keywords : Candida; fluconazole; Antifungal Agents; proteins; electrophoresis; SDS-PAGE.