Abstract

In the present study protein profile of a Candida albicans strain had been examined by chip technology and conventional capillary electrophoresis (CE). Profiles could be characterised by the presence of ten dominating protein peaks. These proteins could be distinguished by both techniques, but their quantity showed significant differences in the electropherograms obtained by CE and chip method. Changes in the protein profile were induced by administration of different antifungal agents. Fluconazole and amphotericin B treatment was able to induce similar changes in the pattern, appearance of a 40-kDa protein and up-regulation of a 60-kDa protein was observed by chip technology. Increase in the quantity of these proteins under stress effect (antifungal treatment) might refer to their stress function in the fungal cell. Treatment of C. albicans cells with MK  94 (fused cyclic Mannich ketone) antifungal compound induced not only the previously mentioned changes, but further specific alterations, appearance of a 19-kDa protein and up-regulation of the low molecular weight proteins. This might refer to the different mode of action of this agent on the fungal cells. Conventional capillary electrophoresis was suitable to detect the appearance of the 19-kDa peak, and up-regulation of the 60 kDa protein, but the other changes could not be detected by this technique. Shorter running time, more effective and baseline separation of proteins refer to the advantages of microchip-based method in the analysis of complex biological samples.

Keywords: Microchip electrophoresis; Capillary electrophoresis; Antifungal drugs; Candida albicans; Stress proteins; Fused cyclic Mannich ketone

Abbreviations: C. albicans, Candida albicans; Hsps, heat shock proteins; MIC, minimum inhibitory concentration; MK, Mannich ketone.

Article Outline

1. Introduction

2. Materials and methods

2.1. Fungal strain and cultivation

2.2. Antimycotics and determination of their minimum inhibitory concentration

2.3. Lysis of fungal cells

2.4. Chip technology

2.5. Capillary electrophoresis

3. Results

3.1. Protein profiles of C. albicans obtained by chip technology

3.2. Protein composition of C. albicans analysed by capillary electrophoresis

4. Discussion

References