Membrane properties of antiviral phospholipids containing heteroatoms in the acyl chains.

Phospholipids containing heteroatoms in the lipid acyl chains, e.g., 1,2-bis(12-methoxydodecanoyl)-sn-3-phosphocholine (L-AC2), exhibit potent anti-HIV activity [Pidgeon, C., Markovich, R. J., Liu, M. D., Holzer, T., Novak, R., & Keyer, K. (1993) J. Biol. Chem. 268, 7773-7778]. AC2 is a synthetic chemical analog of the long-chain phospholipid, dimyristoylphosphatidylcholine (DMPC). Sonicated AC2 lipid dispersions would not entrap either Dextran-4000 or Mn2+ used as aqueous space markers. The lack of entrapment of aqueous space markers indicates that the AC2 structures do not contain an aqueous core that is the characteristic morphology of conventional lipid vesicles formed by sonication. Transmission electron microscopy (TEM) showed that sonicated AC2 lipid dispersions are small homogeneous particles approximately 70-100 A in diameter. 1H NMR experiments using Mn2+ as a broadening reagent indicated that Mn2+ was accessible to all of the AC2 phospholipid headgroups in the AC2 lipid particles formed by sonication. The temperature dependence of 1H spin-lattice (T1) relaxation time measurements revealed that the motional activation energies increased from the choline headgroup to the end of the acyl chains of AC2 molecules in the AC2 lipid particles formed by sonication. Collectively these results demonstrate that AC2 forms micelles. NOESY experiments showed that the AC2 molecules forming the micelle structures have hindered motion compared to conventional short-chain phosphatidylcholine micelles. 31P NMR spectroscopy and TEM showed that the AC2 micelles extensively fuse into giant bilayer liposomes (single-layered) when the temperature is reduced from above to below the main phase transition temperature of AC2. This micelle-to-liposome transition is an irreversible process; increasing the temperature above the Tm does not cause the formation of micelles. Thus, a main finding is that AC2 micelles formed by sonication are not thermodynamically stable because they fuse into large unilamellar vesicles that are stable to further changes in temperature. These unusual membrane properties of sonicated AC2 dispersions may be important for the antiviral activity and metabolism of the phospholipids.