Diacylglycerol and the promotion of lamellar-hexagonal and lamellar-isotropic phase transitions in lipids: Implications for membrane fusion

Changes in steady-state fluorescence anisotropy of 1-(4-trimethylaminophenyl)-6-phenyl-1,3,5-hexatriene (TMA-DPH) are applied to the detection of lamellar-hexagonal transitions in egg phosphatidylethanolamine. Even low (2 mole%) proportions of diacylglycerol decrease the hexagonal transition temperature considerably, as confirmed by differential scanning calorimetry. Diacylglycerol is also found to promote a lamellar to ''isotropic'' (Q(224) cubic) transition in mixtures of phosphatidylcholine:phosphatidylethanolamine:cholesterol. This nonreversible transition is also observed by P-31 nuclear magnetic resonance and detected as a large increase in TMA-DPH steady-state anisotropy. The same technique reveals as well that lysophosphatidylcholine counteracts the effect of diacylglycerol and stabilizes the lamellar phase in both transitions. Diacylglycerol and lysophosphatidylcholine are known to respectively promote and inhibit membrane fusion in a variety of systems. These data are interpreted in support of the hypothesis of a highly bent structural fusion intermediate (''stalk''). They also show the interest of lipid-phase studies in predicting and rationalizing membrane fusion mechanisms.