Daptomycin-Induced Lipid Phases on Model Lipid Bilayers: Effect of Lipid Type and of Lipid Leaflet Order on Membrane Permeability

Daptomycin's bacterial membrane activity is partly due to the defects of lipid-packing at the boundaries of daptomycin-induced, separated lipid phases, which are rich in phosphatidylglycerol (PG). On model membranes, the permeability of phase boundaries is strongly dependent on the extent of saturation of the lipid acyl tails, which affect the lipids' ability to pack within these boundaries, and on the cross-leaflet registration of these boundaries. Using vesicles with asymmetric lipid leaflet compositions, we evaluated the role of headgroup type and/or extent of acyl-tail saturation on daptomycin-induced membrane permeability. We demonstrate that the release rates of vesicle-encapsulated contents scales with the total length of daptomycin-induced, PG-rich phase boundaries. On the outer leaflet, lipids with PG-headgroups (in contact with daptomycin) were a necessary condition, but they still were not adequate for release. Increased membrane permeability was observed only when inner leaflet lipids had saturated acyl tails; we postulate that the latter may have enabled the recruitment, by the outer leaflet daptomycin-induced phases, of inner leaflet lipids in cross-registered phases with boundaries of defective packing that spanned the bilayer. These findings provide insights on the potential role of lipids as a whole (headgroup and acyl tails) and of lipid leaflet order on the boundaries of daptomycin-induced separated lipid phases in model membranes.