Effects of various additives on candidacidal and membrane permeabilizing properties of B4010.

<p>(A) Effect of CCCP and NaN₃ on membrane potential. (B) Effect of various additives on (B) viability and (C) ATP release. (D) Effect of ion-channel inhibitors on membrane potential. The colored arrows indicate the time of addition of additives whereas the black arrows indicate B4010.</p

Interaction of B4010 with model membrane.

<p>(A) Time course of calcein release from SUVs of PC:PE:PS:erg and PC:cholesterol. The peptide:lipid ratio is indicated in the graphs. (B)-(D) Snapshots illustrating the interaction between B4010 and mixed bilayer containing ergosterol. The acyl chains of the aggregated POPC (grey), POPE (cyan), POPS (pink) and ergosterol (orange) are presented in line form. The peptide backbone is shown in ribbon form. (E) Translocation of water molecules (green) from inner leaflet to the outer as a consequence of membrane perturbations caused by B4010.</p

Schematic representation of the peptides used in this study.

<p>The branched lysine is colored in blue. For the scrambled peptide (Sc_B4010) each copy contains the sequence VRGRVRKR.</p

MIC of B4010 against <i>S. cerevisiae</i> mutants carrying altered sterol structure and composition.

a<p>Taken from <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0087730#pone.0087730-TeWelcher1" target="_blank">[29]</a>.</p

Electrostatic potential map on the adsorption of B4010 and B4010_R1A with bilayer.

<p>(A) POPC/POPE/POPS/Erg bilayer (B) B4010-adsorbed bilayer and (C) B4010_R1A-adsorbed bilayer. The negative and positive surfaces are labeled in red and blue, respectively whereas grey color indicates neutral surface.</p

MIC of synthetic linear and branched peptides against various yeasts and fungi.

a<p>Not determined.</p

MIC, cytotoxicity and calcein release properties of tetravalent peptides.

a<p>Effective concentration required to kill 50% of human conjunctival epithelial cells.</p>b<p>Values in parenthesis indicate cell viability in % at the maximum concentration tested.</p>c<p>% calcein released at peptide: liposome ratio 1∶15. n.d. is not determined.</p

Antifungal properties of B4010 in the presence of metal ions and complex biological fluids.

<p>(A) MIC values determined in the presence of monovalent and divalent metal ions. The numbers above the bars indicate the determined MIC values. (B) Candidacidal properties of B4010 in the presence of metal ions. The concentration of B4010 was 5.5 µM. (C) Antifungal activity in the presence of trypsin. (D) Candidacidal activity of B4010 in the presence of 50% rabbit tear fluid.</p

Effect of B4010 on cytoplasmic membrane potential, membrane permeabilization and morphology of <i>C. albicans</i>.

<p>(A) SDS-PAGE showing lack of affinity of B4010 for insoluble chitin. (B) SYTOX Green uptake of <i>C. albicans</i> induced by varying concentration of B4010. (C) B4010-mediated membrane depolarization monitored by diSC₃5 assay. (D) B4010-induced extracellular ATP release of <i>C. albicans</i>. The inset shows the kinetics of ATP release. B4010 concentration was 5.5 µM. (E) SEM of untreated <i>C. albicans</i>. Scale bar is 2 µm (inset scale bar = 200 nm). (F) SEM image of C. albicans treated with 5.5 µM B4010. Scale bar is 1 µm (inset scale bar = 100 nm).</p

Time kill curves for B4010 against <i>C. albicans</i>.

<p>(A) ATCC 10231 and (B) clinical isolate DF2672R. The yeasts cells were incubated with various concentrations of peptide or antifungals in SD broth. At the indicated times, survivors were diluted and plated to allow colony counts.</p