Lateral diffusion and phase separation in two-dimensional solutions of polymerized butadiene lipid in dimyristoylphosphatidylcholine bilayers. A photobleaching and freeze fracture study

Abstract

Mixed vesicles of dimyristoylphosphatidylcholine (DMPC) and a polymerizable lipid containing one diene group per chain are studied by freeze fracture electron microscopy and by the photobleaching (fluorescence recovery after photobleaching) technique. Large thin-walled vesicles of some micron in diameter become more stable after photochemical polymerization. Before polymerization bilayers of the diene lipid exhibit a liquid crystal-to-gel transition at Tg = 31 degrees C. Upon polymerization the transition remains but shifts to a slightly higher temperature (Tg* = 34 degrees C). The transitions in both cases are accompanied by a freezing in of the lateral mobilities. The mixed vesicle exhibits lateral phase separation after polymerization. Before polymerization the two lipids appear miscible at all compositions in the fluid state and at DMPC concentrations at or below 50 mol % in the solid state. After polymerization a two-dimensional solution of the polymer in DMPC is obtained at T greater than Tg*, while lateral phase segregation into DMPC-rich domains and patches of the polymer is observed at T less than Tg*. The domain structure appears identical irrespective of whether polymerization is performed at T greater than Tg or at T less than Tg. A typical value of the diameter of the polymerized lipid domains (approximately 400 A) indicates a rather small aggregation number (N less than 100 monomers). The lateral diffusion coefficient in butadiene-lipid bilayers only decreases from D1 = 3.10(-7) cm2/s to D1 = 8.10(-8) cm2/s (that is by a factor of 4) upon polymerization. This is consistent with the freeze fracture finding of a small aggregation number. We point out the similarities of the mixed vesicles with plasma membranes coupled to the cytoskeleton.