Asymmetry of lipid bilayers induced by monovalent salt: atomistic molecular-dynamics study

Interactions between salt ions and lipid components of biological membranes are essential for the structure, stability, and functions of the membranes. The specific ionic composition of aqueous buffers inside and outside of the cell is known to differ considerably. To model such a situation we perform atomistic molecular-dynamics (MD) simulations of a single-component phosphatidylcholine lipid bilayer which separates two aqueous reservoirs with and without NaCl salt. To implement the difference in electrolyte composition near two membrane sides, a double bilayer setup (i.e., two bilayers in a simulation box) is employed. It turns out that monovalent salt, being in contact with one leaflet only, induces a pronounced asymmetry in the structural, electrostatic, and dynamical properties of bilayer leaflets after 50 ns of MD simulations. Binding of sodium ions to the carbonyl region of the leaflet which is in contact with salt results in the formation of "Na-lipids" complexes and, correspondingly, reduces mobility of lipids of this leaflet. In turn, attractive interactions of chloride ions (mainly located in the aqueous phase close to the water-lipid interface) with choline lipid groups lead to a substantial (more vertical) reorientation of postphatidylcholine headgroups of the leaflet adjoined to salt. The difference in headgroup orientation on two sides of a bilayer, being coupled with salt-induced reorientation of water dipoles, leads to a notable asymmetry in the charge-density profiles and electrostatic potentials of bilayer constitutes of the two leaflets. Although the overall charge density of the bilayer is found to be almost insensitive to the presence of salt, a slight asymmetry in the charge distribution between the two bilayer leaflets results in a nonzero potential difference of about 85 mV between the two water phases. Thus, a transmembrane potential of the order of the membrane potential in a cell can arise without ionic charge imbalance between two aqueous compartments.     

The effect of lipid oxidation on the water permeability of phospholipids bilayers

The effect of lipid oxidation on water permeability of phosphatidylcholine membranes was investigated by means of both scattering stopped flow experiments and atomistic molecular dynamics simulations. Formation of water pores followed by a significant enhancement of water permeability was observed. The molecules of oxidized phospholipids facilitate pore formation and subsequently stabilize water in the membrane interior. A wide range of oxidation ratios, from 15 to 100 mol%, was considered. The degree of oxidation was found to strongly influence the time needed for the opening of a pore. In simulations, the oxidation ratio of 75 mol% was found to be a threshold for spontaneous pore formation in the tens of nanosecond timescale, whereas 15 mol% of oxidation led to significant water permeation in the timescale of seconds. Once a pore was formed, the water permeability was found to be virtually independent of the oxidation ratio.     

Solvent isotope effects on the phase-transition properties of lipid bilayers

Highly sensitive differential scanning microcalorimetry (DSC) has been used to investigate the phase transition properties of lipid vesicles prepared from 1,2-distearoyl-L-3-glyceryl-phosphatidylcholine (DSPC) in H(2)O and D(2)O. The data show that the response of pre-transition properties to D(2)O-->H(2)O substitution is stronger than the main transition properties. We find that there is a small increase in the phase transition temperature (DeltaT approximately 0.5 K) and in the co-operative unit in the main transition. The increase in enthalpy (DeltaH congruent with1 kJ(.)mol(-1)) and in transition temperature (DeltaT congruent with2 K) observed in the pre-transition is comparable with that observed in quite different processes and systems, i.e. melting of nucleic acids and proteins and gel formation. It is suggested that D(2)O-->H(2)O substitution affects the thermal transition in these systems in such a way that the contributions of enthalpy and entropy to structural reorganization of water in these processes is modified.     

A study of reconstituted anion channels using lipid bilayers

A chloride channel from impermeant sarcoplasmic reticulum (SR) was embedded in a planar lipid bilayer (BLM) and its electrical properties determined. Studies using ER-derived vesicles fused with BLMs have shown that they are permeable to Na⁺, K⁺, choline and Cl⁻ but less permeable to Ca²⁺ and Mg²⁺. Though highly permeable to K⁺, the liver ER membrane has been postulated to a lack of an efficient ion-conducting structure for K⁺ channel in the SR. The present study was undertaken with the aim to look at the anionic, Ca²⁺ and K⁺ permeability pathways present in the ER membrane. Our reconstituted system exhibits considerable anionic permeability following the sequence: SCN⁻>I⁻>Br⁻Cl⁻>gluconate. The findings suggest the chloride channels have low field-strength sites. It can be pharmacologically dissected to Zn²⁺-sensitive and DIDS-sensitive types. The gating of the channel is weakly voltage-dependent and at higher positive or negative voltages the channel prefers the low sub-conductance states.     

A simulation study on nanoscale holes generated by gold nanoparticles on negative lipid bilayers

Understanding the interactions of gold nanoparticles (AuNPs) with cellular compartments, especially cell membranes, is of fundamental importance in obtaining their control in biomedical applications. An effort is made in this paper to investigate the interactions of 2.2 nm core AuNPs with negative model bilayer membranes by coarse-grained (CG) molecular dynamics (MD) simulation. The CG model of lipid bilayer was taken from Marrink et al. ( J. Phys. Chem. B 2004, 108, 750-760 ), whereas the CG AuNPs model was developed on the basis of both atomistic MD simulations and experimental data. It was found that AuNPs functionalized with cationic ligands penetrated into the negative bilayer membranes and generated significant disruptions on bilayers. The lipids surrounding the nanoparticle were highly disordered and the bulk surface of the bilayer exhibits some defective areas. Most importantly, it is observed that a nanoscale hole can be formed and expanded spontaneously on the peripheral regions of the 20 × 20 nm bilayer. The expansion of the hole is on the time scale of hundreds of nanosceonds. The fully expanded hole had a radius of ～5.5 nm and could transport water molecules at a rate of up to ～1100 molecule/ns. However holes could not be formed on a larger bilayer (28 × 28 nm). The factors that can eliminate hole formation on the bilayer also include the decrease of cationic lignads on the AuNP, the reduction of negative lipids in the bilayer, the release of bilayer surface tension, the lowering of temperature, and the addition of a high concentration of salt. The results suggest that a hole can only be formed on living cell membranes under extreme conditions.     

Influence of cholesterol on the phase transition of lipid bilayers: a temperature-controlled force spectroscopy study

Cholesterol (Chol) plays the essential function of regulating the physical properties of the cell membrane by controlling the lipid organization and phase behavior and, thus, managing the membrane fluidity and its mechanical strength. Here, we explore the model system DPPC:Chol by means of temperature-controlled atomic force microscopy (AFM) imaging and AFM-based force spectroscopy (AFM-FS) to assess the influence of Chol on the membrane ordering and stability. We analyze the system in a representative range of compositions up to 50 mol % Chol studying the phase evolution upon temperature increase (from room temperature to temperatures high above the T(m) of the DPPC bilayer) and the corresponding (nano)mechanical stability. By this means, we correlate the mechanical behavior and composition with the lateral order of each phase present in the bilayers. We prove that low Chol contents lead to a phase-segregated system, whereas high contents of Chol can give a homogeneous bilayer. In both cases, Chol enhances the mechanical stability of the membrane, and an extraordinarily stable system is observed for equimolar fractions (50 mol % Chol). In addition, even when no thermal transition is detected by the traditional bulk analysis techniques for liposomes with high Chol content (40 and 50 mol %), we demonstrate that temperature-controlled AFM-FS is capable of identifying a thermal transition for the supported lipid bilayers. Finally, our results validate the AFM-FS technique as an ideal platform to differentiate phase coexistence and transitions in lipid bilayers and bridge the gap between the results obtained by traditional methods for bulk analysis, the theoretical predictions, and the behavior of these systems at the nanoscale.     

Thermodynamic study of benzocaine insertion into different lipid bilayers

Despite the general consensus concerning the role played by sodium channels in the molecular mechanism of local anesthetics, the potency of anaesthetic drugs also seems to be related with their solubility in lipid bilayers. In this respect, this work represents a thermodynamic study of benzocaine insertion into lipid bilayers of different compositions by means of molecular dynamics simulation. Thus, the free energy profiles associated with benzocaine insertion into symmetric lipid bilayers composed of different proportions of dipalmitoylphosphatidylcholine and dipalmitoylphosphatidylserine were studied. From the simulation results, a maximum in the free energy (ΔG) profile was measured in the region of the lipid/solution interface. This free energy barrier appears to be very much dependent on the lipid composition of the membrane. On the other hand, the minimum free energy (ΔG) within the bilayer remained almost independent of the lipid composition of the bilayer. By repeating the study at different temperatures, it was seen how the spontaneity of benzocaine insertion into the lipid bilayer is due to an increase in the entropy associated with the process.     

Hydrogen bonded arylamide-linked cholesteryl dimesogenic liquid crystals: a study of the length and side chain effects

Dimesogenic compounds 1a-c, 2a-i, and 3, that are composed of a hydrogen bonding-induced straight arylamide spacer and two appended cholesterol groups, have been designed and synthesized. The backbones of the rigid spacers of 1a-c, 2a-i, and 3 contain one, three, and five benzene units, which bear two, six, and ten alkoxyl (methoxyl, n-octoxyl, or n-dodecoxyl) groups, respectively. The thermal and optical properties of the compounds are investigated by using the differential scanning calorimetry (DSC), polarizing optical microscopy (POM), and powder X-ray diffraction (PXRD) analysis. It is revealed that 1a-c exhibit one or two liquid crystalline (LC) phases, 2a-i exhibit no, one or two LC phases, while 3 exhibits one LC phase in a wide temperature range. Generally, the more and longer alkoxyl chains facilitate the formation of the LC phases at low temperature. Notably, compound 2g, which bears two methoxyl and four dodecoxyl groups, displays a blue-red color change during both the heating and cooling cycle. The result illustrates that dimesogens with large rigid spacers can exhibit different LC phases when long aliphatic chains are appended to balance the strong stacking of the rigid backbones.     

Nonionic urea surfactants: influence of hydrocarbon chain length and positional isomerism on the thermotropic and lyotropic phase behavior

The thermotropic and lyotropic phase behavior of 1- and 5-decyl urea, and 1-, 2-, 4-, and 6-dodecyl urea have been studied. This allowed the effect of positional isomerism to be examined. Intermolecular hydrogen bonding by the urea moiety is the dominant factor in determining the solid-state thermal behavior and crystal solubility boundary of these linear nonionic surfactants. The positional isomers where the urea moiety was not situated at the terminus of the hydrocarbon chain exhibited higher melting points than the 1-alkyl ureas. This has been rationalized by postulating interdigitated chains in the solid state. In the urea surfactant-water systems, three phases are observed, viz. crystalline solid, a dilute aqueous solution of the alkyl urea, and an isotropic liquid. The last two phases coexist in the low-surfactant, high-temperature region of the binary phase diagram. An overview of structure-property correlations for linear nonionic urea surfactants is presented in light of the new physicochemical data obtained for the decyl urea and dodecyl urea positional isomers.     

不等链长的碳氟、碳氢正负离子表面活性剂混合体系的表面活性

研究了全氟辛酸钠与溴化十四烷基三甲铵混合水溶液的表面活性. 测定了不同比例混合物水溶液的表面张力-浓度曲线, 得出临界胶团浓度(cmc)及监 界胶团浓度时的溶液表面张力(γcmc)值. 应用Gibbs吸附公式及吸附层中两表面活性剂分子相互作用参数法求出表面总吸附量、吸附层组成及两表面活性剂分别吸附量等. 指示此吸附层具有多分子层性质. 这可能是碳氢、碳氟正负离子混合体系的特点.     

Interactions of lipid bilayers with supports: a coarse-grained molecular simulation study

The study of lipid structure and phase behavior at the nanoscale is of utmost importance due to implications in understanding the role of the lipids in biochemical membrane processes. Supported lipid bilayers play a key role in understanding real biological systems, but they are vastly underrepresented in computational studies. In this paper, we discuss molecular dynamics simulations of supported lipid bilayers using a coarse-grained model. We first focus on the technical implications of modeling solid supports for biomembrane simulations. We then describe noticeable influences of the support on the systems. We are able to demonstrate that the bilayer system behavior changes when supported by a hydrophilic surface. We find that the thickness of the water layer between the support and the bilayer (the inner-water region in the latter part of this paper) adapts through water permeation on the microsecond time scale. Additionally, we discuss how different surface topologies affect the bilayer. Finally, we point out the differences between the two leaflets induced by the support.     

PH effects on drug interactions with lipid bilayers by liposome electrokinetic chromatography

Liposome electrokinetic chromatography (LEKC) provides convenient and rapid methods for studying drug interactions with lipid bilayers using liposomes as a pseudostationary phase. LEKC was used to determine the effects of pH on the partitioning of basic drugs into liposomes composed of zwitterionic phosphatidylcholine (PC), anionic phosphatidylglycerol (PG), and cholesterol, which mimic the composition of natural cell membranes. An increase in pH results in a smaller degree of ionization of the basic drugs and consequently leads to a lower degree of interaction with the negatively charged membranes. From the LEKC retention data, the fractions of drugs distributed in the bulk aqueous and the liposome phase were determined at various pH values. Finally, lipid mediated shifts in the ionization constants of drugs were examined.     

Bridging across length scales: multi-scale ordering of supported lipid bilayers via lipoprotein self-assembly and surface patterning

We show that a two-step process, involving spontaneous self-assembly of lipids and apolipoproteins and surface patterning, produces single, supported lipid bilayers over two discrete and independently adjustable length scales. Specifically, an aqueous phase incubation of DMPC vesicles with purified apolipoprotein A-I results in the reconstitution of high density lipoprotein (rHDL), wherein nanoscale clusters of single lipid bilayers are corralled by the protein. Adsorption of these discoidal particles to clean hydrophilic glass (or silicon) followed by direct exposure to a spatial pattern of short-wavelength UV radiation directly produces microscopic patterns of nanostructured bilayers. Alternatively, simple incubation of aqueous phase rHDL with a chemically patterned hydrophilic/hydrophobic surface produces a novel compositional pattern, caused by an increased affinity for adsorption onto hydrophilic regions relative to the surrounding hydrophobic regions. Further, by simple chemical denaturation of the boundary protein, nanoscale compartmentalization can be selectively erased, thus producing patterns of laterally fluid, lipid bilayers structured solely at the mesoscopic length scale. Since these aqueous phase microarrays of nanostructured lipid bilayers allow for membrane proteins to be embedded within single nanoscale bilayer compartments, they present a viable means of generating high-density membrane protein arrays. Such a system would permit in-depth elucidation of membrane protein structure-function relationships and the consequences of membrane compartmentalization on lipid dynamics.     

Stretching effects on the permeability of water molecules across a lipid bilayer

Using a coarse grained molecular dynamics model of a solvent-surfactant system, we study the effects of stretching on the permeability of water across a lipid bilayer. The density profile, free energy profile, diffusion profile, and tail ordering parameter were computed for a set of stretched membranes maintained at constant area. We computed the water permeability across each membrane using the inhomogeneous solubility-diffusion model first proposed by Marrink and Berendsen [J. Phys. Chem. 98, 4155 (1994)]. We find that even though the resistance to permeation profile shows a great deal of qualitative change as the membranes are stretched, the overall permeability remains nearly constant within the relevant range of stretching. This is explained by the fact that the main barrier to permeation, located in the densest section of the tails, is insensitive to increased area per lipid, as a result of competing effects. Expansion leads to thinning and a higher density in the tail region, the latter leading to an increase in the free energy barrier. However, this is compensated by the reduction in the transverse distance to cross and a larger diffusion coefficient due to increased disordering in the chains.     

Engineered-membranes: A novel concept for clustering of native lipid bilayers

A strategy for clustering of native lipid membranes is presented. It relies on the formation of complexes between hydrophobic chelators embedded within the lipid bilayer and metal cations in the aqueous phase, capable of binding two (or more) chelators simultaneously Fig. 1. We used this approach with purple membranes containing the light driven proton pump protein bacteriorhodopsin (bR) and showed that patches of purple membranes cluster into mm sized aggregates and that these are stable for months when incubated at 19 °C in the dark. The strategy may be general since four different hydrophobic chelators (1,10-phenanthroline, bathophenanthroline, Phen-C10, and 8-hydroxyquinoline) and various divalent cations (Ni²⁺, Zn²⁺, Cd²⁺, Mn²⁺, and Cu²⁺) induced formation of membrane clusters. Moreover, the absolute requirement for a hydrophobic chelator and the appropriate metal cations was demonstrated with light and atomic force microscopy (AFM); the presence of the metal does not appear to affect the functional state of the protein. The potential utility of the approach as an alternative to assembled lipid bilayers is suggested.     

Surfactant effects of chlorpromazine and imipramine on lipid bilayers containing sphingomyelin and cholesterol

The surface-active drugs chlorpromazine (CPZ) and imipramine (IP) have been tested on large unilamellar vesicles composed of phosphatidylcholine (PC), sphingomyelin (SM), and cholesterol (Ch) in different proportions. The well-characterized nonionic detergent Triton X-100 (TX) has also been used in parallel experiments. Leakage of vesicular aqueous contents and bilayer solubilization have been measured for each surfactant molecule and vesicle composition. All three surface-active molecules behave in a qualitatively similar way, irrespective of bilayer composition: they induce leakage at concentrations well below their critical micellar concentrations (cmc) and solubilization near the cmc. In these events, the potency of the three surfactants under study increases with decreasing cmc, in the order IP相似文献   

Opposing effects of cation binding and hydration on the bending rigidity of anionic lipid bilayers

We correlate the molecularly realistic self-consistent field predictions for the mean bending modulus kc of charged lipid vesicles with experimental observations of the size R of corresponding vesicles that are produced by the freeze-thaw method. We elaborate on the Ansatz that the bending modulus is related to the membrane persistence length and that this length scale sets the radius of the vesicles. Alkali cations have a remarkable effect on the mean bending modulus and thus on the equilibrium radius of negatively charged entropically stabilized dioleoylphosphatidylglycerol (DOPG) vesicles. Where cation hydration typically results in thicker and thus stiffer membranes, specific adsorption to the bilayer surface results in a decrease of the surface charge density and the thickness of the membrane-associated electric double layer. As a result of these opposing effects on kc and R, the largest DOPG vesicles are found in the presence of K+, which combines an intermediate hydration enthalpy and PG-binding affinity.