Emergence of Superstructures from a Homogeneous Lipid Sphere

The spontaneous generation of a periodic hexagonal superstructure on a giant phospholipid sphere (GPS) with a diameter of 20−200 μm was studied. The GPS was composed of ternary phospholipids consisting of dioleoylphosphatidylethanolamine (DOPE), dioleoylphosphatidylcholine (DOPC), and dioleoylphosphatidylinositol-bisphosphate (DOPIP2). GPSs were prepared by natural swelling of a lipid film formed on a glass substrate. A GPS with a homogeneous lipid mixture tends to form a two-layered structure between the surface and inner parts; the surface layer is attributed to a DOPIP2 rich region (we call this layer SL), and the interior is rich in DOPE and DOPC (we call this layer IL). A hexagonal superstructure develops in the SL, and the topology then changes to form multiple-doughnut structures. Finally, myelin-like tubes are generated through symmetry breaking of the doughnutlike structures. The time-dependent change in the surface-area expansion of a GPS is shown to obey the logistic growth model, and this is attributed to the kinetic process of phase segregation between the surface and bulk phase of the GPS

Transformation of ActoHMM Assembly Confined in Cell-Sized Liposome

To construct a simple model of a cellular system equipped with motor proteins, cell-sized giant liposomes encapsulating various amounts of actoHMM, the complexes of actin filaments (F-actin) and heavy meromyosin (HMM, an actin-related molecular motor), with a depletion reagent to mimic the crowding effect of inside of living cell, were prepared. We adapted the methodology of the spontaneous transfer of water-in-oil (W/O) droplets through a phospholipid monolayer into the bulk aqueous phase and successfully prepared stable giant liposomes encapsulating the solution with a physiological salt concentration containing the desired concentrations of actoHMM, which had been almost impossible to obtain using currently adapted methodologies such as natural swelling and electro-formation on an electrode. We then examined the effect of ATP on the cytoskeleton components confined in those cell-sized liposomes, because ATP is known to drive the sliding motion for actoHMM. We added α-hemolysin, a bacterial membrane pore-forming toxin, to the bathing solution and obtained liposomes with the protein pores embedded on the bilayer membrane to allow the transfer of ATP inside the liposomes. We show that, by the ATP supply, the actoHMM bundles inside the liposomes exhibit specific changes in spatial distribution, caused by the active sliding between F-actin and HMM. Interestingly, all F-actins localized around the inner periphery of liposomes smaller than a critical size, whereas in the bulk solution and also in larger liposomes, the actin bundles formed aster-like structures under the same conditions