Fig. 3: Identification of phospholipids as the targets of mandimycin.
From: A polyene macrolide targeting phospholipids in the fungal cell membrane
a,b, MIC-fold changes in mandimycin and amphotericin B against C. albicans BNCC 186382 in the presence of a fixed concentration (1 mg ml−1) and varying concentrations (0.05–1 mg ml−1) of fungal cell membrane components. The assay was performed three times independently (n = 3, two-way ANOVA). c, Schematic illustrating the proposed mode of action of mandimycin on fungal cell membranes. d, ITC assay measuring the binding affinity of mandimycin and amphotericin B to phospholipids and ergosterol. Large unilamellar vesicles (LUVs, 0.1 µm) made by 1,2-dioleoyl-sn-glycero-3-phosphocholine (DOPC) were used as the negative control (n = 3, two-way ANOVA, ****P < 0.0001). e,f, Representative ITC plots (e) and UV–vis spectra (f) of mandimycin and amphotericin B titrated with phosphatidylinositol (PI). The experiments were repeated three times independently, yielding consistent results. g, Knockout of the glycosyltransferase gene mandQ led to the production of the bis-deglycosylated analogue mandimycin B. h,i, Representative ITC plots (h) and UV–vis spectra (i) of mandimycin B titrated with ergosterol and PI. Data are mean ± s.d. in a, b and d. CL, cardiolipin; β-1,3-GLU, β-1,3-glucan; β-1,6-GLU, β-1,6-glucan; Lan, lanosterol; Man, mannose; Mand B, mandimycin B; PC, phosphatidylcholine; PE, phosphatidylethanolamine; PG, phosphatidylglycerol; PS, phosphatidylserine; POC, phosphorylcholine; POE, phosphorylethanolamine; POS, phosphoserine; SM, sphingomyelin. NB, no binding.
