Outside-inside distributions and sizes of mixed phosphatidylcholine-cholesterol vesicles

(1) The effect of the incorporation of cholesterol upon the distribution of various molecular species of phosphatidylcholine across the bilayers of mixed sonicated liposomes (vesicles) has been studied with 31P-MNR. (2) The outside-inside ratio of both saturated and unsaturated phosphatidylcholine species was not much affected by the incorporation of up to 30 mol% cholesterol. Above 30 mol% cholesterol the outside-inside ratio strongly increased for phosphatidylcholines with cis unsaturated fatty acid chains. In contrast the outside-inside ratio of trans unsaturated and fully saturated phophatidylcholine species was either not affected or decreased by the incorporation of more than 30 mol% cholesterol. (3) a simple relationship between the size of the vesicle and the linewidth of the 31P-NMR resonance is described. From the measured linewidths the sizes of the various cholesterol containing vesicles have been obtained. It is found that incorporation of 0-30 mol% cholesterol does not significantly affect the size of the vesicle whereas above 30 mol% cholesterol does not significantly affect the size of the vesicle whereas above 30 mol% cholesterol the size of all phosphatidylcholine vesicles sharply increases. The increase in size is the largest for the more saturated phosphatidylcholine species. (4) From the outside-inside ratio and the size of the vesicle the composition of the outer and inner layer of the mixed vesicles could be obtained. Below 30 mol% cholesterol the composition of outer and inner layer is nearly identical. Above 30 mol% cholesterol the distribution of lipid across the bilayer of all visicles becomes assymetric with a disporportionately larger amount of cholesterol present in the inside monolayer.