Proton-induced fusion of oleic acid-phosphatidylethanolamine liposomes.
作者： Nejat Duzgunes , Robert M. Straubinger , Patricia A. Baldwin , Daniel S. Friend , Demetrios Papahadjopoulos
DOI: 10.1021/BI00334A004
关键词: Phosphatidylserine 、 Phosphatidylethanolamine 、 Phosphatidylcholine 、 Liposome 、 Synthetic membrane 、 Lipid bilayer fusion 、 Chromatography 、 Chemistry 、 Vesicle 、 Oleic acid
摘要: Liposomes composed of oleic acid and phosphatidylethanolamine (3:7 mole ratio) aggregate, become destabilized, fuse below pH 6.5 in 150 mM NaCl. Fusion is monitored by (i) the intermixing internal aqueous contents liposomes, utilizing quenching aminonaphthalene-3,6,8-trisulfonic (ANTS) N,N'-p-xylylenebis(pyridinium bromide) (DPX) encapsulated two separate populations vesicles, (ii) a resonance energy transfer assay for dilution fluorescent phospholipids from labeled to unlabeled (iii) irreversible changes turbidity, (iv) quick-freezing freeze-fracture electron microscopy. Destabilization followed fluorescence increase caused leakage coencapsulated ANTS/DPX or calcein. Ca2+ Mg2+ also induce fusion these vesicles at 3 4 mM, respectively. The threshold higher presence low (subfusogenic) concentrations divalent cations. Vesicles phosphatidylserine/phosphatidylethanolamine acid/phosphatidylcholine do not destabilize, range 7-4, indicating that phosphatidylserine phosphatidylcholine cannot be substituted phosphatidylethanolamine, respectively, proton-induced membrane fusion. Freeze-fracture replicas acid/phosphatidylethanolamine liposomes frozen within 1 s stimulation with 5.3 display larger undergoing fusion, ridges areas bilayer continuity between them. construction pH-sensitive useful as model studying molecular requirements biological systems cytoplasmic delivery macromolecules.
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