The Effect of PS Content on the Ability of Natural Membranes to Fuse with Positively Charged Liposomes and Lipoplexes |
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Authors: | K?Stebelska PM?Dubielecka Email author" target="_blank">AF?SikorskiEmail author |
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Institution: | (1) Laboratory of Cytobiochemistry, Institute of Biochemistry and Molecular Biology, University of Wroclaw, Przybyszewskiego 63/77, Wroclaw, 51-148, Poland;(2) Academic Centre for Biotechnology of Lipid Aggregates, Przybyszewskiego 63/77, Wroclaw, 51-148, Poland |
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Abstract: | Supramolecular aggregates containing cationic lipids have been widely used as transfection mediators due to their ability
to interact with negatively charged DNA molecules and biological membranes. First steps of the process leading to transfection
are partly electrostatic, partly hydrophobic interactions of liposomes/lipoplexes with cell and/or endosomal membrane. Negatively
charged compounds of biological membranes, namely glycolipids, glycoproteins and phosphatidylserine (PS), are responsible
for such events as adsorption, hemifusion, fusion, poration and destabilization of natural membranes upon contact with cationic
liposomes/lipoplexes. The present communication describes the dependence of interaction of cationic liposomes with natural
and artificial membranes on the negative charge of the target membrane, charges which in most cases were generated by charging
the PS content or its exposure. The model for the target membranes were liposomes of variable content of PS or PG (phosphatidylglycerol)
and erythrocyte membranes in which the PS and other anionic compound content/exposure was modified in several ways. Membranes
of increased anionic phospholipid content displayed increased fusion with DOTAP (1,2-dioleoyl-3-trimethylammoniumpropane)
liposomes, while erythrocyte membranes partly depleted of glycocalix, its sialic acid, in particular, showed a decreased fusion
ability. The role of the anionic component is also supported by the fact that erythrocyte membrane inside-out vesicles fused
easily with cationic liposomes. The data obtained on erythrocyte ghosts of normal and disrupted asymmetry, in particular,
those obtained in the presence of Ca2+, indicate the role of lipid flip-flop movement catalyzed by scramblase. The ATP-depletion of erythrocytes also induced an
increased sensitivity to hemoglobin leakage upon interactions with DOTAP liposomes. Calcein leakage from anionic liposomes
incubated with DOTAP liposomes was also dependent on surface charge of the target membranes. In all experiments with the asymmetric
membranes the fusion level markedly increased with an increase of temperature, which supports the role of membrane lipid mobility.
The decrease in positive charge by binding of plasmid DNA and the increase in ionic strength decreased the ability of DOTAP
liposomes/lipoplexes to fuse with erythrocyte ghosts. Lower pH promotes fusion between erythrocyte ghosts and DOTAP liposomes
and lipoplexes. The obtained results indicate that electrostatic interactions together with increased mobility of membrane
lipids and susceptibility to form structures of negative curvature play a major role in the fusion of DOTAP liposomes with
natural and artificial membranes. |
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Keywords: | Cationic liposomes Lipid asymmetry Phosphatidylserine Fusion Gene therapy |
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