Figure 6.

Zinc greatly increases Hst 5 and P113 killing of C. glabrata, and induces almost immediate ATP efflux. C. glabrata Cg931010 cells were used in candidacidal assays and ATP efflux assays. (A) For candidacidal assays, cells were incubated for 1 h at 30 °C with Hst 5 (red), Hst 5ΔMB (blue), and P113 (green) with and without a 1:2 ratio of Zn²⁺ to peptides (hatched) at 7.5 fmol peptides/cell in 10 mM sodium phosphate buffer pH 7.4. Squares and triangles indicate individual replicates. (B) In bioluminescent ATP efflux assays, cells were treated with 0.88 fmol/cell Hst 5 (red), Hst 5ΔMB (blue), or P113 (green) in 10 mM sodium phosphate buffer pH 7.4 with or without added Zn²⁺ at a one to two peptides ratio (hatched). Cells were removed from samples via centrifugation and aliquots of supernatant were taken at 1 and 10 min time points to measure extracellular ATP by luminescence. Both experiments were repeated on at least three separate days and averaged. Significance of differences in killing and ATP efflux were calculated using one way ANOVA with Sidak’s multiple comparison test. * indicates p ≤ 0.05, **** indicates p ≤ 0.0001