Association of the polyene antibiotic amphotericin B with phospholipid vesicles: perturbation by temperature changes

Conformational changes of amphotericin B in the presence of cholesterol as well as in the presence of bilayer vesicles of phosphatidylcholine with saturated fatty acid chains of various lengths (14 less than n less than 22) have been monitored by circular dichroism (CD). It has been shown that the observed species are not only dependent on such parameters as the cholesterol content of the vesicles, the vesicles' physical state, and the number of amphotericin B molecules per vesicle, but also on the time elapsed after mixing and the thermal treatment of the system, which may create irreversible changes. In particular, heating through the transition temperature (Tc) vesicles containing cholesterol and loaded with amphotericin below Tc leads to the expulsion into the aqueous medium of a cholesterol-amphotericin complex, a phenomenon which affords an explanation for some of the electron paramagnetic resonance and resonance Raman results. It has also been shown by gel filtration, ultracentrifugation, and Tc determination that interaction of amphotericin B with vesicles in the gel state induces fusion or aggregation of the vesicles, which is not the case (or at least weakly) when the vesicles are in the liquid crystalline state. This aggregation is the more rapid the nearer the temperature of the reaction is to Tc. This study confirms the great complexity of events which may occur during interaction of amphotericin B with model membranes and presents some results which complement those of studies performed with other spectroscopic methods.