Interaction of the polyene antibiotic amphotericin B with model membranes: differences between small and large unilamellar vesicles.
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 membranes 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 of 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 x 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 Ca2+ 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 x 10(-4)M). In contrast, an important release of the fluorescent dye was observed with DMPC LUV at concentrations of approximately 10(-5)M.(ABSTRACT TRUNCATED AT 250 WORDS)
Milhaud
J
J
Laboratoire de Physique et Chimie Biomoléculaire (CNRS UA198), Université Pierre-et-Marie-Curie, Paris, France.
Hartmann
M A
MA
Bolard
J
J
D000666:Amphotericin B / Q000494:pharmacology*; D001667:Binding, Competitive; D002784:Cholesterol / Q000502:physiology; D002942:Circular Dichroism / Q000379:methods; D004134:Dimyristoylphosphatidylcholine; D004875:Ergosterol / Q000502:physiology; D005452:Fluoresceins; D008567:Membranes, Artificial; D008968:Molecular Conformation; D010539:Permeability
D005452:Fluoresceins; D008567:Membranes, Artificial; C024098:6-carboxyfluorescein; D000666:Amphotericin B; D002784:Cholesterol; D004134:Dimyristoylphosphatidylcholine; D004875:Ergosterol