Transbilayer distribution in small unilamellar phosphatidylglycerol-phosphatidylcholine vesicles

The transbilayer distribution of the phospholipids in small unilamellar vesicles comprised of egg phosphatidylglycerol (PG) and egg phosphatidylcholine (PC) was ascertained by 31P NMR. These vesicles, containing 10-75 mol % PG, were formed by sonication (pH 7.6) and fractionated by centrifugation. Data from spectra accumulated in the presence and absence of a paramagnetic shift reagent, Mn2+, indicated that the phospholipids are randomly arranged across the bilayer. The absence of compositional asymmetry, which contradicts earlier reports, is also exhibited by small unilamellar vesicles (50 mol % PG) prepared by the rapid ethanol injection method. Control experiments showed that Mn2+ did not induce fusion, permeate the vesicles, or cause the phospholipids to migrate across the bilayer. It has been proposed that the transbilayer distribution of charged phospholipids in membranes is a consequence of the different surface charge densities on the opposing sides of the membrane. Our results suggest that it is the difference in the effective polar headgroup volumes of the components rather than the net charge of one component that determines the packing constraints for mixtures of phospholipids with the same acyl chains, at least in highly curved bilayers.