Journal
FREE RADICAL BIOLOGY AND MEDICINE
Volume 52, Issue 11-12, Pages 2302-2311Publisher
ELSEVIER SCIENCE INC
DOI: 10.1016/j.freeradbiomed.2012.03.012
Keywords
Coprisin; Apoptosis; Hydroxyl radicals; Candida albicans; Cytochrome c; Free radicals
Funding
- Next-Generation BioGreen 21 Program, Rural Development Administration, Republic of Korea [PJ008158]
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Coprisin is a 43-mer defensin-like peptide from the dung beetle, Copris tripartitus. Here, we investigated the induction of apoptosis by coprisin in Candida albicans cells. Coprisin exerted antifungal and fungicidal activity without any hemolytic effect. Confocal microscopy indicated that coprisin accumulated in the nucleus of cells. The membrane studies, 1,6-diphenyl-1,3,5-hexatriene, calcein-leakage, and giant unilamellar vesicle assays, confirmed that coprisin did not disrupt the fungal plasma membrane at all. Moreover, the activity of coprisin was energy- and salt-dependent. Next, we investigated whether coprisin induced apoptosis in C. albicans. Annexin V-FITC staining and TUNEL assay showed that coprisin was involved with both the early and the late stages of apoptosis. Coprisin also increased the intracellular reactive oxygen species level, and hydroxyl radicals were included at high levels among the species. The effect of thiourea as a hydroxyl radical scavenger further confirmed the existence of the hydroxyl radicals. Furthermore, coprisin induced mitochondrial membrane potential dysfunction, cytochrome c release, and activation of metacaspases. In summary, this study suggests that coprisin could be a model molecule for a large family of novel antimicrobial peptides possessing apoptotic activity. (C) 2012 Elsevier Inc. All rights reserved.
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