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Tsutomu Hamada

Publications and source records attributed to Tsutomu Hamada.

8 recordsLinked to original sources

Phase diagrams and ordering in charged membranes: Binary mixtures of charged and neutral lipids

We propose a model describing the phase behavior of two-component membranes consisting of binary mixtures of electrically charged and neutral lipids. We take into account the structural phase transition (main-transition) of the hydrocarbon chains, and investigate the interplay between this phase transition and the lateral phase separation. The presence of charged lipids significantly affects the phase behavior of the multi-component membrane. Due to the conservation of lipid molecular volume, the main-transition temperature of charged lipids is lower than that of neutral ones. Furthermore, as compared with binary mixtures of neutral lipids, the membrane phase separation in binary mixtures of charged lipids is suppressed, in accord with recent experiments. We distinguish between two types of charged membranes: mixtures of charged saturated lipid/neutral unsaturated lipid and a second case of mixtures of neutral saturated lipid/charged unsaturated lipid. The corresponding phase behavior is calculated and shown to be very different. Finally, we discuss the effect of added salt on the phase separation and the temperature dependence of the lipid molecular area.

cond-mat.soft

Coupling between pore formation and phase separation in charged lipid membranes

We investigated the effect of charge on the membrane morphology of giant unilamellar vesicles (GUVs) composed of various mixtures containing charged lipids. We observed the membrane morphologies by fluorescent and confocal laser microscopy in lipid mixtures consisting of a neutral unsaturated lipid [dioleoylphosphatidylcholine (DOPC)], a neutral saturated lipid [dipalmitoylphosphatidylcholine (DPPC)], a charged unsaturated lipid [dioleoylphosphatidylglycerol (DOPG$^{\scriptsize{(-)}}$)], a charged saturated lipid [dipalmitoylphosphatidylglycerol (DPPG$^{\scriptsize{(-)}}$)], and cholesterol (Chol). In binary mixtures of neutral DOPC/DPPC and charged DOPC/DPPG$^{\scriptsize{(-)}}$, spherical vesicles were formed. On the other hand, pore formation was often observed with GUVs consisting of DOPG$^{\scriptsize{(-)}}$ and DPPC. In a DPPC/DPPG$^{\scriptsize{(-)}}$/Chol ternary mixture, pore-formed vesicles were also frequently observed. The percentage of pore-formed vesicles increased with the DPPG$^{\scriptsize{(-)}}$ concentration. Moreover, when the head group charges of charged lipids were screened by the addition of salt, pore-formed vesicles were suppressed in both the binary and ternary charged lipid mixtures. We discuss the mechanisms of pore formation in charged lipid mixtures and the relationship between phase separation and the membrane morphology. Finally, we reproduce the results seen in experimental systems by using coarse-grained molecular dynamics simulations.

cond-mat.soft

Charge-induced phase separation in lipid membranes

The phase separation in lipid bilayers that include negatively charged lipids is examined experimentally. We observed phase-separated structures and determined the membrane miscibility temperatures in several binary and ternary lipid mixtures of unsaturated neutral lipid, dioleoylphosphatidylcholine (DOPC), saturated neutral lipid, dipalmitoylphosphatidylcholine (DPPC), unsaturated charged lipid, dioleoylphosphatidylglycerol (DOPG$^{\scriptsize{(-)}}$), saturated charged lipid, dipalmitoylphosphatidylglycerol (DPPG$^{\scriptsize{(-)}}$), and cholesterol. In binary mixtures of saturated and unsaturated charged lipids, the combination of the charged head with the saturation of hydrocarbon tail is a dominant factor for the stability of membrane phase separation. DPPG$^{\scriptsize{(-)}}$ enhances phase separation, while DOPG$^{\scriptsize{(-)}}$ suppresses it. Furthermore, the addition of DPPG$^{\scriptsize{(-)}}$ to a binary mixture of DPPC/cholesterol induces phase separation between DPPG$^{\scriptsize{(-)}}$-rich and cholesterol-rich phases. This indicates that cholesterol localization depends strongly on the electric charge on the hydrophilic head group rather than on the ordering of the hydrocarbon tails. Finally, when DPPG$^{\scriptsize{(-)}}$ was added to a neutral ternary system of DOPC/DPPC/Cholesterol (a conventional model of membrane rafts), a three-phase coexistence was produced. We conclude by discussing some qualitative features of the phase behaviour in charged membranes using a free energy approach.

cond-mat.soft

Micro-Bioreactor Mimicking a Cellular Environment

We report that a cell-sized water droplet (CWD) with a diameter of several tens of microns can serve as a micro-biochemical reactor. Since the droplet inner surface is covered by a phospholipid layer, it provides an environment similar to that in living cells. The CWD is manipulated with laser tweezers and we monitor the time-development of biochemical reactions in a single CWD by fusing two droplets which contain a substrate and an enzyme, respectively. As actual examples of enzymatic reactions, we present results on the reaction of calcein with esterase and on the expression reaction of green fluorescent protein.

physics.bio-ph

Transporting of a Cell-Sized Phospholipid Vesicle Across Water/Oil Interface

When a cell-sized water droplet, with a diameter of several tens of micro meter, is placed in oil containing phospholipids, a stable cell-sized vesicle is spontaneously formed as a water-in-oil phospholipid emulsion (W/O CE) with a phospholipid monolayer. We transferred the lipid vesicle thus formed in the oil phase to the water phase across the water/oil interface by micromanipulation, which suggests that the vesicle is transformed from a phospholipid monolayer as W/O CE into a bilayer. The lipid vesicle can then be transported back into the oil phase. This novel experimental procedure may be a useful tool for creating a model cellular system, which, together with a microreactor, is applicable as a micrometer-scale biochemical reaction field.

physics.chem-ph

Domain Growth Kinetics in a Cell-sized Liposome

We investigated the kinetics of domain growth on liposomes consisting of a ternary mixture (unsaturated phospholipid, saturated phospholipid, and cholesterol) by temperature jump. The domain growth process was monitored by fluorescence microscopy, where the growth was mediated by the fusion of domains through the collision. It was found that an average domain size r develops with time t as r ~ t^0.15, indicating that the power is around a half of the theoretical expectation deduced from a model of Brownian motion on a 2-dimensional membrane. We discuss the mechanism of the experimental scaling behavior by considering the elasticity of the membrane.

cond-mat.soft

Reversible photo-switching in a cell-sized vesicle

A photosensitive amphiphilic molecule can switch the shape of an assembled vesicle as determined by microscopic observation. Photo-isomerization induces a change in membrane fluctuation behavior or a morphological transition between ellipsoid and bud shapes, depending on the asymmetrical degree of the initial shape. The mechanism of this reversible photo-switching in the vesicle morphology is interpreted in terms of a change in the effective cross-sectional area of the photosensitive molecule.

cond-mat.soft

Self Running Droplet: Emergence of Regular Motion from Nonequilibrium Noise

Spontaneous motion of an oil droplet driven by chemical nonequilibricity is reported. It is shown that the droplet undergoes regular rhythmic motion under appropriately designed boundary conditions, whereas it exhibits random motion in an isotropic environment. This study is a novel manifestation on the direct energy transformation of chemical energy into regular spatial-motion under isothermal conditions. A simple mathematical equation including noise reproduces the essential feature of the transition from irregularity into periodic regular motion. Our results will inspire the theoretical study on the mechanism of molecular motors in living matter, working under significant influence of thermal fluctuation.

nlin.AO