Temperature- and pH-controlled fusion between complex lipid membranes. Examples with the diacylphosphatidylcholine/fatty acid mixed liposomes.

The fusion capability of complex lipid bilayers and its pH as well as temperature sensitivity have been studied by optical and spectroscopic means. The aggregation and fusion efficiency of such lipid membranes can be optimized by controlling the phase characteristics of the individual membrane components. For a practically relevant illustration, the stoichiometric 1:2 (mol/mol) mixtures of phosphatidylcholines and fatty acids are used. Perhaps the most interesting liposomes of this kind, which are made of dipalmitoylphosphatidylcholine/elaidic acid (DPPC/ELA-COOH (1:2)), undergo a chain-melting phase transition between 42 degrees C and 48 degrees C, depending on the bulk pH value. The highest chain-melting phase transition temperatures are measured with the fully protonated fatty acids at pH < or = 5.5 and involve a change into the non-bilayer high-temperature state. Upon increasing pH, this transition reverts into an ordinary gel-to-fluid lamellar phase change and occurs at 42 degrees C, by and large. Simultaneously, the rate and the efficacy of fusion between the PC/FA and PC/FA- mixed vesicles decreases. The fusion efficacy of the PC/FA(-) mixed liposomes at pH > or = pK(FA) approximately 7.5 is practically negligible. This is largely due to the increased interbilayer repulsion and to the relatively high water-solubility of the deprotonated fatty acid molecules at high pH. While the pH-variability chiefly affects the efficacy of the intermembrane aggregation, the vesicle fusion itself is more sensitive to temperature variations. It is more likely that the temperature dependence of the intramembrane defect density is chiefly responsible for this. Optimal conditions for the fusion between DPPC/ELA-COOH (1:2) mixed vesicles are thus 3.5 < or = pH < or = 5.5 (6.3) (aggregation maximum) and T > or = 41.5 degrees C = Tm(DPPC) (defect density and fusion maximum). Under such conditions the average size of PC/FA (1:2) mixed vesicles in a 1 mM suspension increases by a factor of 10 over a period of 10 min. Interbilayer fusion can also be catalyzed by the mechanically induced local membrane defects. Freshly made liposomes thus always fuse more avidly than aged vesicles. This permits estimates of the kinetics of membrane defects annihilation based on the measured temporal dependence of the maximum fusion-rate. From such studies, a quasi-exponential decay on the time scale of 1.2 h is found for the thermolabile fusogenic DPPC/ELA-COOH liposomes.