Effects of histatin 5 and derived peptides on Candida albicans.
Three anti-microbial peptides were compared with respect to their killing activity against Candida albicans and their ability to disturb its cellular and internal membranes. Histatin 5 is an anti-fungal peptide occurring naturally in human saliva, while dhvar4 and dhvar5 are variants of its active domain, with increased anti-microbial activity. dhvar4 has increased amphipathicity compared with histatin 5, whereas dhvar5 has amphipathicity comparable with that of histatin 5. All three peptides caused depolarization of the cytoplasmic and/or mitochondrial membrane, indicating membranolytic activity. For the variant peptides both depolarization and killing occurred at a faster rate. With FITC-labelled peptides, no association with the cytoplasmic membrane was observed, contradicting the formation of permanent transmembrane multimeric peptide pores. Instead, the peptides were internalized and act on internal membranes, as demonstrated with mitochondrion- and vacuole-specific markers. In comparison with histatin 5, the variant peptides showed a more destructive effect on mitochondria. Entry of the peptides and subsequent killing were dependent on the metabolic state of the cells. Blocking of the mitochondrial activity led to complete protection against histatin 5 activity, whereas that of dhvar4 was hardly affected and that of dhvar5 was affected only intermediately.
Ruissen
A L
AL
Academic Centre for Dentistry Amsterdam (ACTA), Department of Dental Basic Sciences, Section of Oral Biochemistry, Van der Boechorststraat 7, 1081 BT Amsterdam, The Netherlands. ala.ruissen.obc.acta@med.vu.nl
Groenink
J
J
Helmerhorst
E J
EJ
Walgreen-Weterings
E
E
Van't Hof
W
W
Veerman
E C
EC
Nieuw Amerongen
A V
AV
D000595:Amino Acid Sequence; D000935:Antifungal Agents / Q000737:chemistry / Q000493:pharmacokinetics / Q000494:pharmacology*; D002176:Candida albicans / Q000187:drug effects* / Q000378:metabolism; D016650:Fluorescein-5-isothiocyanate; D005456:Fluorescent Dyes; D014644:Genetic Variation; D054808:Histatins; D008564:Membrane Potentials / Q000187:drug effects; D018613:Microscopy, Confocal; D008969:Molecular Sequence Data; D012471:Salivary Proteins and Peptides / Q000737:chemistry / Q000235:genetics / Q000494:pharmacology*; D013347:Subcellular Fractions / Q000378:metabolism; D014617:Vacuoles / Q000187:drug effects / Q000378:metabolism
D000935:Antifungal Agents; D005456:Fluorescent Dyes; C068075:HTN3 protein, human; D054808:Histatins; D012471:Salivary Proteins and Peptides; D016650:Fluorescein-5-isothiocyanate