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Antonie Van Leeuwenhoek. 2001 Sep;79(3-4):297-309.

Killing of Candida albicans by histatin 5: cellular uptake and energy requirement.

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  • 1Department of Periodontology and Oral Biology, Boston University Goldman School of Dental Medicine, Massachusetts 02118-2392, USA.

Abstract

Histatins, a group of histidine-rich proteins in human saliva, exhibit antimicrobial activity and are therefore considered to be important in the prevention of infections in the oral cavity. Although killing of C. albicans by histatins has been extensively studied, little is known about the processes responsible for this antifungal activity. Recent studies show the requirement of metabolic activity and ATP production for histatin 5 killing activity. Therefore, the goal of this study was to investigate the kinetics of histatin 5 interaction at different temperatures with C. albicans wild type cells and with respiratory deficient mutants of C. albicans. Synthetic histatin 5 was labeled with fluorescein-5-isothiocyanate (FITC) and its association with C. albicans cells was followed by epi-fluorescence microscopy and fluorescence confocal microscopy. At 37 degrees C, histatin 5 accumulates intracellularly, and both killing activity and uptake of unlabeled and FITC-labeled histatin 5 are time- and concentration-dependent. At 4 degrees C, no killing is observed and FITC-histatin 5 is only associated with the cytoplasmic membrane. Internalization and killing activity only occurs after cells are transferred to 37 degrees C. In addition, cellular accumulation of histatin 5 is concomitant with a moderate alteration of membrane integrity leading to the release of UV-absorbing cell components into the medium. The uptake of histatin 5, the release of UV-absorbing materials and killing of C. albicans are markedly decreased by the respiratory inhibitor sodium azide. Concomitantly, respiratory deficient mutants of C. albicans are also less susceptible to histatin 5. These results indicated that histatin 5 killing activity could be directly correlated to histatin 5 internalization. Both of these processes are prevented by modulators of cellular metabolic activity.

PMID:
11816973
[PubMed - indexed for MEDLINE]
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