title: Candidacidal mechanism of a Leu/Lys-rich α-helical amphipathic model antimicrobial peptide and its diastereomer composed of D,L-amino acids.
abstract: We investigated the mechanism of candidacidal action of a Lys/Leu-rich α-helical model antimicrobial peptide (K(9)L(8)W) and its diastereomeric peptide (D(9)-K(9)L(8)W) composed of D,L-amino acids. K(9)L(8)W killed completely Candida albicans within 30 min, but D(9)-K(9)L(8)W killed only 72% of C. albicans even after 100 min. Tryptophan fluorescence spectroscopy indicated that the fungal cell selectivity of D(9)-K(9)L(8)W is closely correlated with a selective interaction with the negatively charged PC/PE/PI/ergosterol (5:2.5:2.5:1, w/w/w/w) phospholipids, which mimic the outer leaflet of the plasma membrane of C. albicans. K(9)L(8)W was able to induce almost 100% calcein leakage from PC/PE/PI/ergosterol (5:2.5:2.5:1, w/w/w/w) liposomes at a peptide:lipid molar ratio of 1:16, whereas D(9)-K(9)L(8)W caused only 25% dye leakage even at a peptide:lipid molar ratio of 1:2. Confocal laser-scanning microscopy revealed that FITC-labeled D(9)-K(9)L(8)W penetrated the cell wall and cell membrane and accumulated inside the cells, whereas FITC-labeled K(9)L(8)W did not penetrate but associated with the membranes. Collectively, our results demonstrated that the candidacidal activity of K(9)L(8) W and D(9)-K(9)L(8)W may be due to the transmembrane pore/channel formation or perturbation of the fungal cytoplasmic membranes and the inhibition of intracellular functions, respectively. Finally, D(9)-K(9)L(8)W with potent anti-Candida activity but no hemolytic activity may be potentially a useful lead compound for the development of novel antifungal agents.
authors.affiliation: Department of Bio-Materials, Graduate School, Chosun University, Gwangju 501-759, Republic of Korea.
authors.forename: Peng
authors.initials: P
authors.lastname: Wang
authors.forename: Yong Hai
authors.initials: YH
authors.lastname: Nan
authors.forename: Song Yub
authors.initials: SY
authors.lastname: Shin
mesh_terms: D000595:Amino Acid Sequence; D000935:Antifungal Agents / Q000494:pharmacology*; D023181:Antimicrobial Cationic Peptides / Q000737:chemistry / Q000494:pharmacology*; D002176:Candida albicans / Q000187:drug effects*; D002462:Cell Membrane / Q000187:drug effects; D008969:Molecular Sequence Data; D010743:Phospholipids / Q000737:chemistry; D017433:Protein Structure, Secondary; D013237:Stereoisomerism
chemical_list: D000935:Antifungal Agents; D023181:Antimicrobial Cationic Peptides; C552028:K9L8W peptide; D010743:Phospholipids