PHOTOACTIVATED ENHANCEMENT OF LIPOSOME FUSION

The photopolymerization of two-component large unilamellar Liposomes (LUV) composed of 3:1 dioleoylphosphatidylethanolamine (DOPE) and either 1,2-bis[10-(2'-hexadienoyloxy)decanoyl]-sn-glycero-3-phosphatidylcholine (bis-SorbPC) or 1-palmitoyl-2-[10-(2'-hexadienoyloxy)decanoyl]-sn-glycero-3-phosphatidylcholine (mono-SorbPC) facilitated liposome fusion. Fusion was characterized by fluorescent assays for lipid mixing, aqueous contents mixing, and aqueous contents leakage. The rate and extent of the liposome fusion was dependent on the extent of photopolymerization, temperature, and the fusion initiation conditions, including the pH and the presence of Mg2+ ions. Examination of the temperature dependence of fusion for unpolymerized and polymerized liposomes showed that an enhancement of the rate of fusion occurred in the temperature range Delta T-I, which previous NMR studies have identified as the initial appearance of precursors to the formation of the inverted cubic phase [Barry, J. A., et al. (1992) Biochemistry 31, 10114]. The phase behavior and fusion characteristics of the DOPE/bis-SorbPC (3:1) membranes provide unequivocal evidence that liposome fusion is mediated via intermediates associated with the lamellar to Q(II) phase transition rather than the H-II phase, Photopolymerization of SorbPC-containing liposomes forms poly-SorbPC, which enhances the lateral separation of the liposome components. The formation of enriched domains of polymorphic lipids, e.g., DOPE, causes isothermal induction of fusion by lowering the critical fusion temperature of the membranes.