Kinetics of cholesterol and phospholipid exchange between mycoplasma membranes and lipid vesicles

The following modifications in the lipid composition of Mycoplasma gallisepticum or M. capricolum membranes led to changes in the rates of [14C]cholesterol and [14C]phospholipid exchange between cell membranes and an excess of small unilamellar vesicles: 1) increase in the cholesterol/phospholipid molar ratio from 0.25 to 0.92; 2) incorporation of synthetic, cross-linked phosphatidylethanolamine (PE) derivatives, 3) incorporation of sphingomyelin (SPM); and 4) increase in the phosphatidylglycerol/cardiolipin ratio of M. capricolum cell membranes by supplementing the growth medium with 0.5 mM CaCl2 CaCl2, and decrease in this ratio by supplementing the growth medium with 0.5 mM CaCl2 and 20 micrograms/ml egg phosphatidylcholine or with isopalmitic acid. The rates of radiolabeled lipid exchange decreased when the content of cholesterol, cross-linked PE, or SPM increased, indicating an inverse correlation between exchange rate and membrane lipid order. This is consistent with an exchange mechanism that involves the slow dissolution of the lipid from the surface of the donor particle. Lipidic particles appeared in trypsin-treated M. capricolum membranes obtained from cells grown with both Ca2+ and PC, whereas the hexagonal-II phase appeared in membranes from cells grown with Ca2+. Cholesterol and phospholipid exchange rates were enhanced under the conditions in which the bilayer structure was destabilized by transitional states between the lamellar and hexagonal-II phases. Thus, mycoplasmas are well suited for examination of the influence of membrane lipid composition on rates of lipid exchange between membranes.