Purification and comparison of heat shock protein 90 (Hsp90) in Candida albicans isolates from Malaysian and Iranian patients and infected mice

Objective: The purposes of this study were to purify and compare the concentration ratios of heat shock protein 90 (Hsp90) in clinical isolates of Candida albicans (C. albicans) obtained from Malaysian and Iranian patients and infected mice. Materials and methods: Hsp90 was extracted using glass beads and ultracentrifugation from yeast cells and purified by ion exchange chromatography (DEAE-cellulose) and followed by affinity chromatography (hydroxyapatite). Purity of Hsp90 was controlled by SDS-PAGE and its identification was realized by immunoblotting test. Results: The graphs of ion exchange and affinity chromatography showed one peak in all C. albicans isolates obtained from both Malaysian and Iranian samples, infected mice and under high-thermal (42 °C) and low-thermal (25 °C) shock. In immunoblotting, the location of Hsp90 fragments was obtained around 47, 75 and 82 kDa. The least average concentration ratios of Hsp90 were 0.350 and 0.240 mg/g for Malaysian and Iranian isolates at 25 °C, respectively, while the highest average concentration ratios of Hsp90 were 3.05 and 2.600 mg/g for Malaysian and Iranian isolates at 42 °C, respectively. There were differences in the ratio amount of Hsp90 between Malaysian isolates (1.01 ± 0.07 mg/g) and mice kidneys (1.23 ± 0.28 mg/g) as well as between Iranian isolates (0.70 ± 0.19 mg/g) and mice kidneys (1.00 ± 0.28 mg/g) (P < 0.05). Conclusion: The results showed differences in all situations tested including Iranian and Malaysian isolates, samples treated with temperatures (25 °C or 42 °C) and before and after infecting the mice (37 °C), indicating higher virulent nature of this yeast species in high temperature in human and animal models

Differential gene expression of heat shock protein 90 (Hsp90) of Candida albicans obtained from Malaysian and Iranian patients

Background: Candida albicans (C. albicans) has several virulence factors, in particular heat shock protein 90 (Hsp90), which is expressed by Hsp90 gene. The purposes of this study were to assess the expression of Hsp90 gene in clinical and control isolates of C. albicans obtained from different geographical regions (Malaysia and Iran), different temperatures (25°C, 37°C and 42°C) and mice with candidiasis. Methods: C. albicans isolates were cultured onto sabouraud dextrose agar (SDA). The assessment of the expression of Hsp90 gene was performed using real time-polymerase chain reaction (RT-PCR). Results: The results showed a significant increase in the expression of C. albicans Hsp90 gene under high thermal shock (42°C) when compared to other temperatures tested (P-value = 0.001). The mean differences in the expression of Hsp90 gene at 37°C were 0.20 (95% confidence interval (CI) 0.13-0.29) between Malaysian and Iranian controls (P-value = 0.040) and 0.47 (95% CI 0.27-0.60) between Malaysian and Iranian patients (P-value = 0.040). Conclusion: The results demonstrated that the expression of C. albicans Hsp90 gene varied between Malaysian and Iranian subjects, representing the efficacy of geographical and thermal conditions on virulence gene expression