Membrane Elastic Properties Research Articles

Asymmetric membrane "sticky tape" enables simultaneous relaxation of area and curvature in simulation.

Biological lipid membranes are generally asymmetric, not only with respect to the composition of the two membrane leaflets but also with respect to the state of mechanical stress on the two sides. Computer simulations of such asymmetric membranes pose unique challenges with respect to the choice of boundary conditions and ensemble in which such simulations are to be carried out. Here, we demonstrate an alternative to the usual choice of fully periodic boundary conditions: The membrane is only periodic in one direction, with free edges running parallel to the single direction of periodicity. In order to maintain bilayer asymmetry under these conditions, nanoscale "sticky tapes" are adhered to the membrane edges in order to prevent lipid flip-flop across the otherwise open edge. In such semi-periodic simulations, the bilayer is free to choose both its area and mean curvature, allowing for minimization of the bilayer elastic free energy. We implement these principles in a highly coarse-grained model and show how even the simplest examples of such simulations can reveal useful membrane elastic properties, such as the location of the monolayer neutral surface.

Numerical study on honeycomb membrane-type acoustic metamaterial with added mass

This paper presents numerical investigations on the transmission loss (TL) of a honeycomb structure with embedded membrane to which small masses are attached. The structure is a lightweight honeycomb membrane-type acoustic metamaterial that presents excellent transmission loss especially at low frequencies. The added mass is a solid material that is attached to the membrane, which is modelled as a linear isotropic elastic material with fixed boundary conditions. The influences of the membrane material properties and of the added mass parameters on the transmission loss are presented. It is demonstrated that the resonant peak amplitude of the transmission loss and its frequency band can be controlled by the properties of the added mass. The various numerical simulation results of configurations with and without added masses as well as various membrane elastic properties are compared. The honeycomb structure used in this study constitutes generally the core layer in sandwich honeycomb panels. While the TL of the honeycomb structure alone is zero, due to the open cell distribution, the integration of a membrane within the honeycomb with added masses induces a significant improvement of the transmission loss. This lightweight metamaterial is shown to attenuate the low frequency noise.

Triggering the Amphotericin B Pore-Forming Activity by Phytochemicals.

The macrolide polyene antibiotic amphotericin B (AmB), remains a valuable drug to treat systemic mycoses due to its wide antifungal activity and low probability of developing resistance. The high toxicity of AmB, expressed in nephropathy and hemolysis, could be partially resolved by lowering therapeutic AmB concentration while maintaining efficacy. This work discusses the possibility of using plant polyphenols and alkaloids to enhance the pore-forming and consequently antifungal activity of AmB. We demonstrated that phloretin, phlorizin, naringenin, taxifolin, quercetin, biochanin A, genistein, resveratrol, and quinine led to an increase in the integral AmB-induced transmembrane current in the bilayers composed of palmitoyloleoylphosphocholine and ergosterol, while catechin, colchicine, and dihydrocapsaicin did not practically change the AmB activity. Cardamonin, 4'-hydroxychalcone, licochalcone A, butein, curcumin, and piperine inhibited AmB-induced transmembrane current. Absorbance spectroscopy revealed no changes in AmB membrane concentration with phloretin addition. A possible explanation of the potentiation is related to the phytochemical-produced changes in the elastic membrane properties and the decrease in the energy of formation of the lipid mouth of AmB pores, which is partially confirmed by differential scanning microcalorimetry. The possibility of AmB interaction with cholesterol in the mammalian cell membranes instead of ergosterol in fungal membranes, determines its high toxicity. The replacement of ergosterol with cholesterol in the membrane lipid composition led to a complete loss or a significant decrease in the potentiating effects of tested phytochemicals, indicating low potential toxicity of these compounds and high therapeutic potential of their combinations with the antibiotic. The discovered combinations of AmB with plant molecules that enhance its pore-forming ability in ergosterol-enriched membranes, seem to be promising for further drug development in terms of the toxicity decrease and efficacy improvement.

Nanoscale membrane dynamics in chain asymmetric lipid bilayers.

Lipids that have two tails of different lengths are found throughout biomembranes in nature, yet the effects of this asymmetry on the membrane properties are not well understood, especially the membrane dynamics. Accommodating such a difference in tail length could significantly impact the membrane dynamics and elastic properties depending on how the longer tail is situated within the bilayer. To probe these effects, here we study the nanoscale bending fluctuations in model chain-asymmetric 14:0-18:0 PC (MSPC) and 18:0-14:0 PC (SMPC) lipid bilayers using neutron spin echo (NSE) spectroscopy. We find that despite the partial interdigitation that is known to persist in the fluid phase of these membranes, the collective fluctuations are enhanced on timescales of tens of nanoseconds, and the chain-asymmetric lipid bilayers are softer than an analogous chain-symmetric lipid with the same average number of carbons in the acyl tails, di-16:0 PC (DPPC). Quantitative comparison of the NSE results suggests that the enhanced bending fluctuations at the nanosecond timescales are consistent with experimental and computational studies that showed the compressibility moduli of chain-asymmetric lipid membranes are 20 % to 40% lower than chain-symmetric lipid membranes. These studies add to growing evidence that the partial interdigitation in chain-asymmetric lipid membranes in the fluid phase is highly dynamic and influences membrane dynamic processes from the molecular to mesoscopic length scales without significantly changing the bilayer thickness or area per lipid.

Investigating structure-property relations in cholesterol-rich lipid membranes.

Cholesterol is a key molecule in eukaryotic cell membranes. It mediates essential membrane processes, including viral budding and the mechanical resistance of membranes against pathogens. Hence, understanding the influence of cholesterol on the mechanics of functional membranes is crucial. Cholesterol is known to cause lipids to pack more tightly, manifesting in a decrease in the average area per lipid, AL, at the aqueous interface. Yet, how variations in packing density influence the elastic properties of lipid membranes with varying degrees of chain unsaturation remains largely unexplored. Theories based on mean field calculations have predicted various forms of the interdependence between the bending rigidity modulus, κ, and lipid packing density. However, direct assessments of these predictions have been limited, due to the difficulty in accessing the membrane's mechanical properties on the nanoscale. To address this, we performed studies using neutron spin-echo spectroscopy, solid-state 2H NMR relaxometry, and molecular dynamics simulations on lipid membranes with different degrees of chain unsaturation. We find that cholesterol has a stiffening effect on all studied membranes over the accessible length and time scales. Our findings show that κ has an inverse dependence on AL, indicating that cholesterol controls membrane elastic properties by modifying lipid packing density. As AL decreases, the number of conformations accessible to the lipid chains also decreases and the membrane becomes more rigid. Yet, different expressions of κ vs. AL are obtained when we consider reduced variables. Establishing well-defined structure-property relationships will open up new possibilities for predicting the behavior of more complex biological membranes and will inform design rules for engineered membranes and artificial cells with real-world functionalities.

Janus magnetoelastic membrane swimmers.

Soft swimming microrobots have attracted considerable attention due to their potential applications in diverse fields ranging from biomedicines to environmental remediation. The locomotion control is of importance to the research of micromachines and microrobots. Inspired by the motility strategies of living microorganisms, such as flagella, cilia, and euglenoids, we focus on propulsion mechanisms with a design of Janus magnetoelastic crystalline membrane microswimmers actuated by time-varying magnetic fields. Such a Janus swimmer consists of a ferromagnetic cap completed by a magnetoelastic membrane body, where superparamagnetic particles are uniformly distributed on the surface. Under the influence of external magnetic fields, the swimmer undergoes complex shape transitions due to the interplay between the magnetic dipole-dipole interactions, the elasticity of the magnetoelastic membranes, and also the hydrodynamics of surrounding fluids. We show that those shape changes are nonreciprocal, which can generate locomotion such that the propulsion speed can be optimized by tailoring the membrane elastic properties. Besides, we also demonstrate that the Janus swimmer can be magnetically guided in a spiral trajectory. With such adequate control of locomotion in both speed and direction via non-invasive magnetic fields, this study provides another promising candidate design for the future development of microswimmers.

Molecular simulations and NMR reveal how lipid fluctuations affect membrane mechanics

Lipid bilayers form the main matrix of functional cell membranes, and their dynamics underlie a host of physical and biological processes. Here we show that elastic membrane properties and collective molecular dynamics (MD) are related by the mean-square amplitudes (order parameters) and relaxation rates (correlation times) of lipid acyl chain motions. We performed all-atom MD simulations of liquid-crystalline bilayers that allow direct comparison with carbon-hydrogen (CH) bond relaxations measured with NMR spectroscopy. Previous computational and theoretical approaches have assumed isotropic relaxation, which yields inaccurate description of lipid chain dynamics and incorrect data interpretation. Instead, the new framework includes a fixed bilayer normal (director axis) and restricted anisotropic motion of the CH bonds in accord with their segmental order parameters, enabling robust validation of lipid force fields. Simulated spectral densities of thermally excited CH bond fluctuations exhibited well-defined spin-lattice (Zeeman) relaxations analogous to those in NMR measurements. Their frequency signature could be fit to a simple power-law function, indicative of nematic-like collective dynamics. Moreover, calculated relaxation rates scaled as the squared order parameters yielding an apparent κC modulus for bilayer bending. Our results show a strong correlation with κC values obtained from solid-state NMR studies of bilayers without and with cholesterol as validated by neutron spin-echo measurements of membrane elasticity. The simulations uncover a critical role of interleaflet coupling in membrane mechanics and thus provide important insights into molecular sites of emerging elastic properties within lipid bilayers.

PEO-b-PBD Diblock Copolymers Induce Packing Defects in Lipid/Hybrid Membranes and Improve Insertion Rates of Natively Folded Peptides.

Hybrid membranes assembled from biological lipids and synthetic polymers are a promising scaffold for the reconstitution and utilization of membrane proteins. Recent observations indicate that inclusion of small fractions of polymer in lipid membranes can improve protein folding and function, but the exact structural and physical changes a given polymer sequence imparts on a membrane often remain unclear. Here, we use all-atom molecular dynamics simulations to study the structure of hybrid membranes assembled from DOPC phospholipids and PEO-b-PBD diblock copolymers. We verified our computational model using new and existing experimental data and obtained a detailed picture of the polymer conformations in the lipid membrane that we can relate to changes in membrane elastic properties. We find that inclusion of low polymer fractions induces transient packing defects into the membrane. These packing defects act as insertion sites for two model peptides, and in this way, small amounts of polymer content in lipid membranes can lead to large increases in peptide insertion rates. Additionally, we report the peptide conformational space in both pure lipid and hybrid membranes. Both membranes support similar alpha helical peptide structures, exemplifying the biocompatibility of hybrid membranes.

Amyloid β Dodecamer Disrupts the Neuronal Membrane More Strongly than the Mature Fibril: Understanding the Role of Oligomers in Neurotoxicity.

The amyloid cascade hypothesis states that senile plaques, composed of amyloid β (Aβ) fibrils, play a key role in Alzheimer’s disease (AD). However, recent experiments have shown that Aβ oligomers are more toxic to neurons than highly ordered fibrils. The molecular mechanism underlying this observation remains largely unknown. One of the possible scenarios for neurotoxicity is that Aβ peptides create pores in the lipid membrane that allow Ca2+ ions to enter cells, resulting in a signal of cell apoptosis. Hence, one might think that oligomers are more toxic due to their higher ability to create ion channels than fibrils. In this work, we study the effect of Aβ42 dodecamer and fibrils on a neuronal membrane, which is similar to that observed in AD patients, using all-atom molecular dynamics simulations. Due to short simulation times, we cannot observe the formation of pores, but useful insight on the early events of this process has been obtained. Namely, we showed that dodecamer distorts the lipid membrane to a greater extent than fibrils, which may indicate that ion channels can be more easily formed in the presence of oligomers. Based on this result, we anticipate that oligomers are more toxic than mature fibrils, as observed experimentally. Moreover, the Aβ–membrane interaction was found to be governed by the repulsive electrostatic interaction between Aβ and the ganglioside GM1 lipid. We calculated the bending and compressibility modulus of the membrane in the absence of Aβ and obtained good agreement with the experiment. We predict that the dodecamer will increase the compressibility modulus but has little effect on the bending modulus. Due to the weak interaction with the membrane, fibrils insignificantly change the membrane elastic properties.

Modeling the mesoscopic consequences of specific interactions on biomembranes

The continuum description of lipid membranes via the Helfrich functional and its extensions is widely employed in the modeling of membranes at the mesoscopic scale. A major problem in the application of this theory to membrane remodeling by proteins and other adsorbates is the local modification of membrane shape, lipid composition and elastic properties by adsorbates, such as proteins. We address this problem by direct incorporation of specific interactions into the local parametrization of the continuum model.

Membrane-MEDYAN: Simulating Deformable Vesicles Containing Complex Cytoskeletal Networks.

The plasma membrane defines the shape of the cell and plays an indispensable role in bridging intra- and extracellular environments. Mechanochemical interactions between plasma membrane and cytoskeleton are vital for cell biomechanics and mechanosensing. A computational model that comprehensively captures the complex, cell-scale cytoskeleton-membrane dynamics is still lacking. In this work, we introduce a triangulated membrane model that accounts for the membrane's elastic properties, as well as for membrane-filament steric interactions. The corresponding force-field was incorporated into the active biological matter simulation platform, MEDYAN ("mechanochemical dynamics of active networks"). Simulations using the new model shed light on how actin filament bundling affects generation of tubular membrane protrusions. In particular, we used membrane-MEDYAN simulations to investigate protrusion initiation and dynamics while varying geometries of filament bundles, membrane rigidities and local G-Actin concentrations. We found that the bundles' protrusion propensities sensitively depend on the synergy between bundle thickness and inclination angle at which the bundle approaches the membrane. The new model paves the way for simulations of biological systems involving intricate membrane-cytoskeleton interactions, such as those occurring at the leading edge and the cortex, eventually helping to uncover the fundamental principles underlying the active matter organization in the vicinity of the membrane.

Does membrane curvature elastic energy play a role in mediating oxidative stress in lipid membranes?

The effects of oxidative stress on cells are associated with a wide range of pathologies. Oxidative stress is predominantly initiated by the action of reactive oxygen species and/or lipoxygenases on polyunsaturated fatty acid containing lipids. The downstream products are oxidised phospholipids, bioactive aldehydes and a range of Schiff base by-products between aldehydes and lipids, or other biomacromolecules. In this review we assess the impact of oxidative stress on lipid membranes, focusing on the changes that occur to the curvature preference (lipid spontaneous curvature) and elastic properties of membranes, since these biophysical properties modulate phospholipid homeostasis. Studies show that the lipid products of oxidative stress reduce stored curvature elastic energy in membranes. Based upon this observation, we hypothesize that the effects of oxidative stress on lipid membranes will be reduced by compounds that increase stored curvature elastic energy. We find a strong correlation appears across literature studies that we have reviewed, such that many compounds like vitamin E, Curcumin, Coenzyme Q10 and vitamin A show behaviour consistent with this hypothesis. Finally, we consider whether age-related changes in lipid composition represent the homeostatic response of cells to compensate for the accumulation of in vivo lipid oxidation products.

A microfluidic methodology to identify the mechanical properties of capsules: comparison with a microrheometric approach

We present a microfluidic method to measure the elastic properties of a population of microcapsules (liquid drops enclosed by a thin hyperelastic membrane). The method is based on the observation of flowing capsules in a cylindrical capillary tube and an automatic inverse analysis of the deformed profiles. The latter requires results from a full numerical model of the fluid–structure interaction accounting for nonlinear membrane elastic properties. For ease of use, we provide them under the form of databases, when the initially spherical capsule has a membrane governed by a neo-Hookean or a general Hooke's law with different surface Poisson ratios. Ultimately, the microfluidic method yields information on the type of elastic constitutive law that governs the capsule wall material together with the value of the elastic parameters. The method is applied to a population of ovalbumin microcapsules and is validated by means of independent experiments of the same capsules subjected to a different flow in a microrheological device. This is of great interest for quality control purposes, as small samples of capsule suspensions can be diverted to a measuring test section and mechanically tested with a 10 % precision using an automated process.

Elastic moduli of lipid membranes: Reproducibility of AFM measures

Membrane elastic properties play a major role in membrane remodeling events, such as vesicle fusion and fission. They are also crucial in drug delivery by liposomes. Different experimental techniques are available to measure elastic properties. Among them, atomic force microscopy (AFM) presents the unique advantage of being directly applicable to nano-sized liposomes. Unfortunately, different AFM measures reported in the literature show little agreement among each other and are difficult to compare with measures of bending modulus obtained by other experimental techniques or by molecular simulations. In this work we determine the bending rigidity of Egg PC liposomes in terms of Young modulus via AFM measurements, using two different tip shapes and different cantilever force constants. We interpret the measures using the Hertz and Shell models, and observe a clear dependency of the Young modulus values on the tip properties and on the interpretative theory. The effect of the AFM tip shape is less important than the effect of the cantilever force constant, and the mathematical model has a major effect on the interpretation of the data. The Shell theory provides the closest agreement between AFM data and other experimental data for the membrane bending modulus. Finally, we compare the results to calculations of bending modulus from molecular dynamics simulations of membrane buckles. Simulations provide values of bending modulus consistent with literature data, but the agreement with AFM experiments is reasonable only for some specific experimental conditions.

Membrane Elastic Properties During Neural Precursor Cell Differentiation.

Neural precursor cells differentiate into several cell types that display distinct functions. However, little is known about how cell surface mechanics vary during the differentiation process. Here, by precisely measuring membrane tension and bending modulus, we map their variations and correlate them with changes in neural precursor cell morphology along their distinct differentiation fates. Both cells maintained in culture as neural precursors as well as those plated in neurobasal medium reveal a decrease in membrane tension over the first hours of culture followed by stabilization, with no change in bending modulus. During astrocyte differentiation, membrane tension initially decreases and then increases after 72 h, accompanied by consolidation of glial fibrillary acidic protein expression and striking actin reorganization, while bending modulus increases following observed alterations. For oligodendrocytes, the changes in membrane tension are less abrupt over the first hours, but their values subsequently decrease, correlating with a shift from oligodendrocyte marker O4 to myelin basic protein expressions and a remarkable actin reorganization, while bending modulus remains constant. Oligodendrocytes at later differentiation stages show membrane vesicles with similar membrane tension but higher bending modulus as compared to the cell surface. Altogether, our results display an entire spectrum of how membrane elastic properties are varying, thus contributing to a better understanding of neural differentiation from a mechanobiological perspective.

The Effect of Ions on Membrane Elasticity - Implications for Vesicle Fusion

Ions and charged peptides can have dramatic effects on the properties of membranes. A case in point is calcium and its crucial role in vesicle fusion.[1] Using all-atom molecular dynamics, we simulated calcium mediated vesicle fusion.[2] Aided by a new, local approach to determining membrane elastic properties,[3] we are able to quantify the effect of ions on lipid spontaneous curvature, bending rigidity, and tilt. We probe the effect of ions on membranes of different composition. Our simulations indicate that calcium specifically induces negative spontaneous curvature in those anionic lipid membranes which are susceptible to calcium mediated fusion, concomitantly strongly affecting the Gaussian bending modulus. We further determine the mechanism of membrane curvature induction through ion-headgroup interactions, giving special attention to lipid clustering. In addition to membrane elasticity, we discuss electrostatic effects on osmotic pressures and the hydration force. This systematic approach allows us to fill crucial gaps in our understanding of the mechanism of membrane fusion. Finally, we point out possible implications for synaptic vesicle fusion and endocytosis. [1] E. Neher, R. Schneggenburger, Nature 406, 889-893 (2000) [2] C. Allolio, A. Magarkar, P. Jurkiewicz, K. Baxova, M. Javanainen, R. Šachl, M. Hof, D. Horinek, V. Heinz, R. Rachel, C. Ziegler, M. Cebecauer, A. Schrofel, P. Jungwirth, resubmitted to PNAS (2018) [3] C. Allolio, A. Haluts, D. Harries,Chem. Phys, in press (2018)

Atomic force microscopy reveals new biophysical markers for monitoring subcellular changes in oxidative injury: Neuroprotective effects of quercetin at the nanoscale.

Oxidative stress has been recognised as an important pathological mechanism underlying the development of neurodegenerative diseases. The biomarkers for assessing the degree of oxidative stress have been attracting much interest because of their potential clinical relevance in understanding the cellular effects of free radicals and evaluation of the efficacy of drug treatment. Here, an interdisciplinary approach using atomic force microscopy (AFM) and cellular and biological molecular methods were used to investigate oxidative damage in P19 neurons and to reveal the underlying mechanism of protective action of quercetin. Biological methods demonstrated the oxidative damage of P19 neurons and showed that quercetin improved neuronal survival by preventing H2O2-induced p53 and Bcl-2 down-regulation and modulated Akt and ERK1/2 signalling pathways. For the first time, AFM was employed to evaluate morphologically (roughness, height, Feret dimension) and nanomechanical (elasticity) properties in H2O2-induced neuronal damage. The AFM analysis revealed that quercetin suppressed H2O2-provoked changes in cell membrane elasticity and morphological properties, thus confirming its neuroprotective activity. The obtained results indicate the potential of AFM-measured parameters as a biophysical markers of oxidative stress-induced neurodegeneration. In general, our study suggests that AFM can be used as a highly valuable tool in other biomedical applications aimed at screening and monitoring of drug-induced effects at cellular level.

Membrane Elastic Deformations Modulate Gramicidin A Transbilayer Dimerization and Lateral Clustering

Gramicidin A (gA) is a short β-helical peptide known to form conducting channels in lipid membranes because of transbilayer dimerization. The gA conducting dimer, being shorter than the lipid bilayer thickness, deforms the membrane in its vicinity, and the bilayer elastic energy contributes to the gA dimer formation energy. Likewise, membrane incorporation of a gA monomer, which is shorter than the lipid monolayer thickness, creates a void, thereby forcing surrounding lipid molecules to tilt to fill it. The energy of membrane deformation was calculated in the framework of the continuum elasticity theory, taking into account splay, tilt, lateral stretching/compression, Gaussian splay deformations, and external membrane tension. We obtained the interaction energy profiles for two gA monomers located either in the same or in the opposite monolayers. The profiles demonstrated the long-range attraction and short-range repulsion behavior of the monomers resulting from the membrane deformation. Analysis of the profile features revealed conditions under which clusters of gA monomers would not dissipate because of diffusion. The calculated dependence of the dimer formation and decay energy barriers on the membrane elastic properties was in good agreement with the available experimental data and suggested an explanation for a hitherto contentious phenomenon.

Determining the Bending Moduli of Asymmetric Bilayers by Simulation

Biological membranes are almost always asymmetric in terms of lipid composition. Such asymmetry is thought to play an important part in biological functions via alteration of the membrane's elastic properties, including its bending modulus. Recently experimental techniques have been developed which enable the preparation of asymmetric GUVs and therefore allow the measurement and direct comparison of bending moduli for pure bilayers, mixed symmetric bilayers and asymmetric bilayers composed of similar lipid species. These experiments show that asymmetry does indeed have a marked effect on bilayer stiffness. In order to reproduce these data and develop a molecular level understanding of them, we have taken a computational approach using both atomistic and coarse grained forcefields for a variety of system sizes and lipid species. We have performed analysis of our trajectories using a range of theoretical models based on a variety of different analysis techniques including those based directly on membrane area compressibility; those based on spectral analysis involving 2D discrete Fourier transforms; and those based on the distribution of angles of splay. Our results show that the different models employed provide consistent results across a range of system sizes both with each other and the experimental data, as well as suggesting promising new directions for experimental investigation.

Cell membrane biophysics with optical tweezers.

Membrane elastic properties play important roles in regulating cell shape, motility, division and differentiation. Here I review optical tweezer (OT) investigations of membrane surface tension and bending modulus, emphasizing didactic aspects and insights provided for cell biology. OT measurements employ membrane-attached microspheres to extract long cylindrical nanotubes named tethers. The Helfrich-Canham theory yields elastic parameters in terms of tether radius and equilibrium extraction force. It assumes initial point-like microsphere attachment and no cytoskeleton content within tethers. Experimental force-displacement curves reveal violations of those assumptions, and I discuss proposed explanations of such discrepancies, as well as recommended OT protocols. Measurements of elastic parameters for predominant cell types in the central nervous system yield correlations between their values and cell function. Micro-rheology OT experiments extend these correlations to viscoelastic parameters. The results agree with a quasi-universal phenomenological scaling law and are interpreted in terms of the soft glass rheology model. Spontaneously-generated cell nanotube protrusions are also briefly reviewed, emphasizing common features with tethers. Filopodia as well as tunneling nanotubes (TNT), which connect distant cells and allow transfers between their cytoplasms, are discussed, including OT tether pulling from TNTs which mediate communication among bacteria, even of different species. Pathogens, including bacteria, viruses and prions, opportunistically exploit TNTs for cell-to-cell transmission of infection, indicating that TNTs have an ancient evolutionary origin.