The creation of effective fungal membrane models for neutron and X-ray reflectometry experiments is a key step in the development of new antifungal pharmaceuticals and agrochemicals to allow in vitro investigation of their mode of interaction with target cells. The structure of the obtained models depends on the properties of the lipids used and the final composition of the leaflets, and can be subject to the spontaneous translocation of phospholipids across the bilayer. The effect of phospholipid acyl-chain unsaturation and the presence of steroids in the membrane on the bilayer asymmetry were examined by means of neutron reflectometry. The measurements showed that membrane stability was higher if a zwitterionic, saturated acyl-chain phospholipid is present as the inner leaflet. Furthermore, membrane asymmetry was higher in the case of fully saturated lipid systems. As a result, membrane models consisting of fully saturated acyl chains within the inner leaflet are recommended as the starting point for subsequent studies of antifungal interactions owing to the simplicity of the models and their relative stability, thus allowing better control over the exact lipid composition facing the tested antifungal.
Keywords: fungal membranes; lipid bilayers; neutron reflectometry.