We investigated the phase separation of dioleoylphosphatidylserine (DOPS) and dipalmitoylphosphatidylcholine (DPPC) in giant unilamellar vesicles in hypotonic solution using fluorescence and confocal laser scanning microscopy. Although phase separation in charged lipid membranes is generally suppressed by the electrostatic repulsion between the charged headgroups, osmotic stress can promote the formation of charged lipid domains. Interestingly, we observed three-phase coexistence even in DOPS/DPPC binary lipid mixtures. The three phases were DPPC-rich, dissociated DOPS-rich, and nondissociated DOPS-rich phases. The two forms of DOPS were found to coexist owing to the ionization of the DOPS headgroup, such that the system could be regarded as quasi-ternary. The three formed phases with differently ionized DOPS domains were successfully identified experimentally by monitoring the adsorption of positively charged particles. In addition, coarse-grained molecular dynamics simulations confirmed the stability of the three-phase coexistence. Attraction mediated by hydrogen bonding between protonated DOPS molecules and reduction of the electrostatic interactions at the domain boundaries stabilized the three-phase coexistence.
The response of lipid bilayers to osmotic stress is an important part of cellular function. Previously, in [Oglecka et al. 2014], we reported that cell-sized giant unilamellar vesicles (GUVs) exposed to hypotonic media, respond to the osmotic assault by undergoing a cyclical sequence of swelling and bursting events, coupled to the membrane's compositional degrees of freedom. Here, we seek to deepen our quantitative understanding of the essential pulsatile behavior of GUVs under hypotonic conditions, by advancing a comprehensive theoretical model for vesicle dynamics. The model quantitatively c…
By means of lattice-based Monte Carlo simulations, we address properties of two-component lipid membranes on the experimentally relevant spatial scales of order of a micrometer and time intervals of order of a second, using DMPC/DSPC lipid mixtures as a model system. Our large-scale simulations allowed us to obtain important results previously not reported in simulation studies of lipid membranes. We find that, within a certain range of lipid compositions, the phase transition from the fluid phase to the fluid-gel phase coexistence proceeds via near-critical fluctuations, while for other lipid…
We report here on the pitfalls of the simulation model introduced in the "Capturing Phase Behavior of Ternary Lipid Mixtures with a Refined Martini Coarse-Grained Force Field" [Journal of Chemical Theory and Computation 2018, 14, 11, 6050-6062]. This refined Martini model was reported to reproduce experimental phase diagrams for a ternary DOPC/DPPC/cholesterol mixture, including the coexistence of two liquid phases. However, we demonstrate that this coexistence only emerged due to an unfortunate choice of simulation parameters, which leads to poor energy conservation. Specifically, the constra…
A question of considerable interest to cell membrane biology is whether phase segregated domains across an asymmetric bilayer are strongly correlated with each other and whether phase segregation in one leaflet can induce segregation in the other. We answer both these questions in the affirmative, using an atomistic molecular dynamics simulation to study the equilibrium statistical properties of a 3-component {\em asymmetric} lipid bilayer comprising an unsaturated POPC (palmitoyl-oleoyl-phosphatidyl-choline), a saturated SM (sphingomyelin) and cholesterol with different composition ratios. Ou…
We devise a soft, solvent-free, coarse-grained model for lipid bilayer membranes. The non-bonded interactions take the form of a weighted-density functional which allows us to describe the thermodynamics of self-assembly and packing effects of the coarse-grained beads in terms of a density expansion of the equation of state and the weighting functions that regularize the microscopic bead densities, respectively. Identifying the length and energy scales via the bilayer thickness and the thermal energy scale, kT, the model qualitatively reproduces key characteristics (e.g., bending rigidity, are…
We investigate theoretically the behavior of proteins as well as other large macromolecules which are incorporated into amphiphilic monolayers at the air-water interface. We assume the monolayer to be in the coexistence region of the ``main'' transition, where domains of the liquid condensed phase coexist with the liquid expanded background. Using a simple mean-field free energy accounting for the interactions between proteins and amphiphilic molecules, we obtain the spatial protein distribution with the following characteristics. When the proteins preferentially interact with either the liqui…
Understanding the (de)mixing behavior of multicomponent lipid bilayers is an important step towards unraveling the nature of spatial composition heterogeneities in cellular membranes and their role in biological function. We use coarse-grained molecular dynamics simulations to study the composition phase diagram of a quaternary mixture of phospholipids and cholesterol. This mixture is known to exhibit both uniform and coexisting phases. We compare and combine different statistical measures of membrane structure to identify the onset of phase coexistence in composition space. An important eleme…
Using binary hard disk mixture models, we studied the two-dimensional ordering of bacteriorhodopsins in a lipid bilayer. The phase diagrams were calculated using the thermodynamic perturbation theory. We examined two types of effective interactions to discuss the lateral depletion effects caused by repulsive core interaction between lipid molecules. The results indicate that the core repulsions drastically broaden the coexistence region for the fluid--ordered phase.
We propose a model describing liquid-solid phase coexistence in mixed lipid membranes by including explicitly the occurrence of a rippled phase. For a single component membrane, we employ a previous model in which the membrane thickness is used as an order parameter. As function of temperature, this model properly accounts for the phase behavior of the three possible membrane phases: solid, liquid and the rippled phase. Our primary aim is to explore extensions of this model to binary lipid mixtures by considering the composition dependence of important model parameters. The obtained phase diag…
We introduce a simple and predictive model for determining the phase stability of ternary phospholipid-cholesterol mixtures. Assuming that competition between the liquid and gel order of the phospholipids is the main driving force behind lipid segregation, we derive a Gibbs free-energy of mixing, based on the thermodynamic properties of the lipids main transition. A numerical approach was devised that enable the fast and efficient determination of the ternary diagrams associated with our Gibbs free-energy. The computed phase coexistence diagram of DOPC/DPPC/cholesterol reproduces well known fe…
Though 1-alkanols are well known to have anesthetic properties, the mode of operation remains enigmatic. We perform extensive atomistic molecular dynamics simulation to study the partitioning of 1- alkanols of different chain-lengths in raft-like model membrane made up of mixture of unsaturated dioleoyl-phosphatidylcholine (DOPC) and saturated dipalmitoyl-phosphatidylcholine (DPPC) and choles- terol (Chol) exhibiting phase coexistence of liquid-ordered (lo) - liquid disordered (ld) phase domains. Our simulation shows that the effect of 1-alkanols on the mechanical properties of the membrane ha…
A great variety of biologically relevant monolayers present phase coexistence characterized by domains formed by lipids in an ordered phase state dispersed in a continuous, disordered phase. The difference in surface densities between these phases originates inter-domain dipolar interactions, which are relevant for the determination of the spacial distribution of domains, as well as their dynamics. In this work, we propose a novel manner of estimating the dipolar repulsion using a passive method that involves the analysis of images of the monolayer with phase coexistence. The method is based o…
In spite of the great success that all-atom molecular dynamics simulations have seen in revealing the nature of the lipid bilayer, the interplay between a membrane's curvature and dynamics remains elusive. This is largely due to the computational challenges involved in simulating a highly curved membrane, as the one found in a small vesicle. In the present work, thanks to the computing power of Anton2, we present the first all-atom molecular dynamics simulation of a full-scale, realistically composed (both heterogeneous and asymmetric) vesicle of a meaningful time scale (over 10 microseconds),…
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 uns…
We propose a model that accounts for the budding transition of asymmetric two-component lipid domains, where the two monolayers (leaflets) have different average compositions controlled by independent chemical potentials. Assuming a coupling between the local curvature and local lipid composition in each of the leaflets, we discuss the morphology and thermodynamic behavior of asymmetric lipid domains. The membrane free-energy contains three contributions: the bending energy, the line tension, and a Landau free-energy for a lateral phase separation. Within a mean-field treatment, we obtain vari…
We characterize the order-disorder transition in a model lipid bilayer using molecular dynamics simulations. We find that the ordered phase is hexatic. In particular, in-plane structures possess a finite concentration of 5-7 disclination pairs that diffuse throughout the plane of the bilayer, and further, in-plane structures exhibit long-range orientational order and short-range translational order. In contrast, the disordered phase is liquid. The transition between the two phases is first order. Specifically, it exhibits hysteresis, and coexistence exhibits an interface with capillary scaling…
Stripe-like domains of Langmuir monolayers formed by surfactants with partially fluorinated lipid anchors (F-alkyl lipids) are observed at the gas-liquid phase coexistence. The average periodicity of the stripes, measured by fluorescence microscopy, is in the micrometer range, varying between 2 and 8 microns. The observed stripe-like patterns are stabilized due to dipole-dipole interactions between terminal -CF3 groups. These interactions are particularly strong as compared with non-fluorinated lipids due to the low dielectric constant of the surrounding media (air). These long-range dipolar i…
Mixtures of lipids and cholesterol are commonly used as model systems for studying the formation of liquid-ordered ($L_o$) domains in heterogeneous biological membranes. The simplest model system exhibiting coexistence between $L_o$ domains and a liquid-disordered ($L_d$) matrix is that of a binary mixture of saturated lipids like DPPC and cholesterol (Chol). DPPC/Chol mixtures have been investigated for decades both experimentally, theoretically, and recently also by means of atomistic simulations. Here, we present a minimal lattice model that captures the correct behavior of this mixture acr…
We consider the phase separation in an asymmetrically charged lipid bilayer membrane consisting of neutral and negatively charged lipids that are in contact with in/out ionic solutions having different ionic strengths. The two asymmetric leaflets are coupled through electrostatic interactions. Based on a free energy approach, the critical point and phase diagrams are calculated for different ionic strength of the two solutions and coupling parameter. An increase of the coupling constant or asymmetry in the salt concentration between the in/out solutions yields a higher phase separation tempera…
Despite a vast clinical application of anesthetics, the molecular level of understanding of general anesthesia is far from our reach. Using atomistic molecular dynamics simulation, we study the effects of common anesthetics: ethanol, chloroform and methanol in the fully hydrated symmetric multicomponent lipid bilayer membrane comprising of an unsaturated palmitoyl-oleoyl-phosphatidyl-choline (POPC), a saturated palmitoyl-sphingomyelin (PSM) and cholesterol (Chol) which exhibits phase coexistence of liquid-ordered (lo) - liquid disordered (ld) phase domains. We find that the mechanical and phys…
We study the stability and shapes of domains with spontaneous curvature in fluid films and membranes, embedded in a surrounding membrane with zero spontaneous curvature. These domains can result from the inclusion of an impurity in a fluid membrane, or from phase separation within the membrane. We show that for small but finite line and surface tensions and for finite spontaneous curvatures, an equilibrium phase of protruding circular domains is obtained at low impurity concentrations. At higher concentrations, we predict a transition from circular domains, or "caplets", to stripes. In both ca…
We have studied the collective short wavelength dynamics in deuterated DMPC bilayers by inelastic neutron scattering. The corresponding dispersion relation $\hbarω$(Q) is presented for the gel and fluid phase of this model system. The temperature dependence of the inelastic excitations indicates a phase coexistence between the two phases over a broad range and leads to a different assignment of excitations than that reported in a preceding inelastic x-ray scattering study [Phys. Rev. Lett. {\bf 86}, 740 (2001)]. As a consequence, we find that the minimum in the dispersion relation is actually …
Monolayer spontaneous curvatures for cholesterol, DOPE, POPE, DOPC, DPPC, DSPC, POPC, SOPC, and egg sphingomyelin were obtained using small-angle X-ray scattering (SAXS) on inverted hexagonal phases (HII). Spontaneous curvatures of bilayer forming lipids were estimated by adding controlled amounts to a HII forming template following previously established protocols. Spontanous curvatures of both phosphatidylethanolamines and cholesterol were found to be at least a factor of two more negative than those of phosphatidylcholines, whose J0 are closer to zero. Interestingly, a significant positive …
The study of the structural behaviour of pure and multi-component lipids at various temperatures and the interaction of these multi-component lipids with pharmaceutically important drugs carry huge importance. Here, we investigated the phase behaviour of the pure PSPC (1-palmitoyl-2-stearoyl-sn-glycero-3-phosphocholine), and multicomponent PSPC and DSPE-PEG2000(1,2-distearoyl-sn-glycero-3-phosphoethanolamine-N[amino(polyethylene glycol)-2000]) membranes at seven different temperatures ranging from 280 K to 360 K, and calculated their structural properties. We observe a transition from the gel …
Multicomponent lipid mixtures exhibit complex phase behavior, including coexistence of nanoscopic fluid phases in ternary mixtures mimicking the composition of the outer leaflet of mammalian plasma membrane. The physical mechanisms responsible for the small size of phase domains are unknown, due in part to the difficulty of determining the size and lifetime distributions of small, fleeting domains. Steady-state FRET provides information about the spatial distribution of lipid fluorophores in a membrane, and with an appropriate model can be used to determine the size of phase domains. Starting …
We propose a model that accounts for budding behavior of domains in lipid bilayers, where each of the bilayer leaflets has a coupling between its local curvature and local lipid composition. The compositional asymmetry between the two monolayers leads to an overall spontaneous curvature. The membrane free-energy contains three contributions: bending energy, line tension, and a Landau free-energy for a lateral phase separation. Within a mean-field treatment, we obtain various phase diagrams which contain fully-budded, dimpled and flat states. In particular, for some range of membrane parameters…
Multi-component membranes such as ternary mixtures of lipids and cholesterol can exhibit coexistence regions between two liquid phases. When such membranes adhere to a corrugated substrate, the phase separation process strongly depends on the interplay between substrate topography, bending rigidities, and line tension of the membrane domains as we show theoretically via energy minimization and Monte Carlo simulations. For sufficiently large bending rigidity contrast between the two membrane phases, the corrugated substrate truncates the phase separation process and leads to a stable pattern of…
We report on the modeling of the dynamics of confined lipid membranes. We derive a thin film model in the lubrication limit which describes an inextensible liquid membrane with bending rigidity confined between two adhesive walls. The resulting equations share similarities with the Swift-Hohenberg model. However, inextensibility is enforced by a time-dependent nonlocal tension. Depending on the excess membrane area available in the system, three different dynamical regimes, denoted as A, B and C, are found from the numerical solution of the model. In regime A, membranes with small excess area …
