Interaction of the polyene antibiotic amphotericin B with model membranes: differences between small and large unilamellar vesicles

Abstract

The interaction of the polyene antibiotic amphotericin B (AmB) (Fig. 1) with large unilamellar vesicles (LUV) was monitored by circular dichroism (CD) and carboxyfluorescein (CF) release. LUV afford a far better model for biological membrannes than small unilamellar vesicles (SUV) which have been used until now. With dimyristoyl phosphatidyl choline (DMPC) LUV (i.e., containing saturated acyl chains), a strong and not saturable binding for AmB/lipid ratios up to 0.5 was observed both above and below the phase transition temperature. Incorporation to cholesterol into the vesicles did not significantly change the interaction. With egg PC (EPC) LUV (i.e., containing unsaturated acyl chains), quite a different picture emerged: the binding reached saturation for AmB/lipid ratios of about 5 ×10 −3, a result not observed with EPC SUV. When sterols were introduced into membranes, the CD spectral features obtained in the presence of ergosterol were different from those obtained in the presence of cholesterol. Such a different behavior was not observed with SUV. We suggest that species whose CD spectrum was observed after 15 min in the presence of ergosterol-containing EPC LUV is the particular one which forms wide channels and induces a Ca 2+ release. (H. Ramos, A. Attias, B.E. Cohen and J. Bolard, submitted for publication). The CF release from EPC LUV induced by AmB was very low, even at very high concentrations of the antibiotic (3 × 10 −4M). In contrast, an important release of the fluorescent dye was observed with DMPC LUV at concentrations of ≈ 10 −5M. These results are compared with recent data concerning the interaction of LUV with the pantaenic antibiotic filipin (Fig. 1) (J. Milhaud, J. Mazerski, E. Dufourc and J. Bolard, submitted; J. Milhaud, J. Bolard, P. Benveniste and M.A. Hartmann [1988] Biochim. Biophys. Acta 943, 315–325).