Abstract
Lipid membrane asymmetry plays an important role in cell function and activity, being for instance a relevant signal of its integrity. The development of artificial asymmetric membranes thus represents a key challenge. In this context, an emulsion‐centrifugation method is developed to prepare giant vesicles with an asymmetric membrane composed of an inner monolayer of poly(butadiene)‐b‐poly(ethylene oxide) (PBut‐b‐PEO) and outer monolayer of 1‐palmitoyl‐2‐oleoyl‐sn‐glycero‐3‐phosphocholine (POPC). The formation of a complete membrane asymmetry is demonstrated and its stability with time is followed by measuring lipid transverse diffusion. From fluorescence spectroscopy measurements, the lipid half‐life is estimated to be 7.5 h. Using fluorescence recovery after photobleaching technique, the diffusion coefficient of 1,2‐dioleoyl‐sn‐glycero‐3‐phosphoethanolamine‐N‐(lissamine rhodamine B sulfonyl) (DOPE‐rhod, inserted into the POPC leaflet) is determined to be about D = 1.8 ± 0.50 μm2 s−1 at 25 °C and D = 2.3 ± 0.7 μm2 s−1 at 37 °C, between the characteristic values of pure POPC and pure polymer giant vesicles and in good agreement with the diffusion of lipids in a variety of biological membranes. These results demonstrate the ability to prepare a cell‐like model system that displays an asymmetric membrane with transverse and translational diffusion properties similar to that of biological cells.
Highlights
Choline derivatives such as phosphatidylcholine or sphingomyelin are exposed in higher proportions on theThe mimicking and reproduction of natural components or external monolayer
A previously reported emulsion-centrifugation protocol[58] was adapted to afford asymmetric giant hybrid unilamellar vesicles with an asymmetric giant hybrid polymer–lipid unilamellar vesicles (aGHUV) consisting of an inner leaflet of poly(butadiene)b-poly(ethylene oxide) (PBut2.5-b-PEO1.3) and an outer leaflet of 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine (POPC) lipid
With help of centrifugation and the difference of density between sucrose and glucose, the polymer-stabilized sucrose droplets cross the interface while getting coated with an outer monolayer of lipid, and the resulting POPC/PBut2.5b-PEO1.3 aGHUV can be recovered in the lower glucose phase (Figure 1a)
Summary
Choline derivatives such as phosphatidylcholine or sphingomyelin are exposed in higher proportions on theThe mimicking and reproduction of natural components or external monolayer. Lecommandoux activities between both sides of the membrane and results
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