DOPC Bilayers Research Articles

Studying the Effects of Dissolved Noble Gases and High Hydrostatic Pressure on the Spherical DOPC Bilayer Using Molecular Dynamic Simulations.

Fine-grained molecular dynamics simulations have been conducted to depict lipid objects enclosed in water and interacting with a series of noble gases dissolved in the medium. The simple point-charge (SPC) water system, featuring a boundary composed of 1,2-Dioleoyl-sn-glycero-3-phosphocholine (DOPC) molecules, maintained stability throughout the simulation under standard conditions. This allowed for the accurate modeling of the effects of hydrostatic pressure at an ambient pressure of 25 bar. The chosen pressure references the 240 m depth of seawater: the horizon frequently used by commercial divers, who comprise the primary patient population of the neurological complication of inert gas narcosis and the consequences of high-pressure neurological syndrome. To quantify and validate the neurological effects of noble gases and discriminate them from high hydrostatic pressure, we reduced the dissolved gas molar concentration to 1.5%, three times smaller than what we previously tested for the planar bilayer (3.5%). The nucleation and growth of xenon, argon and neon nanobubbles proved consistent with the data from the planar bilayer simulations. On the other hand, hyperbaric helium induces only a residual distorting effect on the liposome, with no significant condensed gas fraction observed within the hydrophobic core. The bubbles were distributed over a large volume-both in the bulk solvent and in the lipid phase-thereby causing substantial membrane distortion. This finding serves as evidence of the validity of the multisite distortion hypothesis for the neurological effect of inert gases at high pressure.

Concentration-Dependent Effects of Curcumin on Membrane Permeability and Structure.

Growing evidence suggests that many bioactive molecules can nonspecifically modulate the physicochemical properties of membranes and influence the action of embedded membrane proteins. This study investigates the interactions of curcumin with protein-free model membranes consisting of 1,2-dioleoyl-sn-glycero-3-phosphocholine (DOPC) and DOPC with cholesterol (4/1 mol ratio). The focus is on the capability of curcumin to modify membrane barrier properties such as water permeability assayed through the droplet interface bilayer (DIB) model membrane. For pure DOPC, our findings show a concentration-dependent biphasic effect: a reduction in water permeability is observed at low concentrations (up to 2 mol %), whereas at high concentrations of curcumin, water permeability increases. In the presence of cholesterol, we observed an overall reduction in water permeability. A combination of complementary experimental methods, including phase transition parameters studied by differential scanning calorimetry (DSC) and structural properties measured by attenuated total reflectance (ATR)-FTIR, provides a deeper understanding of concentration-dependent interactions of curcumin with DOPC bilayers in the absence and presence of cholesterol. Our experimental findings align with a molecular mechanism of curcumin's interaction with model membranes, wherein its effect is contingent on its concentration. At low concentrations, curcumin binds to the lipid-water interface through hydrogen bonding with the phosphate headgroup, thereby obstructing the transport of water molecules. Conversely, at high concentrations, curcumin permeates the acyl chain region, inducing packing disorders and demonstrating evidence of phase separation. Enhanced knowledge of the impact of curcumin on membranes, which, in turn, can affect protein function, is likely to be beneficial for the successful translation of curcumin into effective medicine.

Differential Effects of Soy Isoflavones on the Biophysical Properties of Model Membranes.

The effects that the main soy isoflavones, genistein and daidzein, have upon the biophysical properties of a model lipid bilayer composed of 1,2-dioleoyl-sn-glycero-3-phosphocholine (DOPC) or DOPC with cholesterol (4 to 1 mol ratio) have been investigated by transbilayer water permeability, differential scanning calorimetry, and confocal Raman microspectroscopy. Genistein is found to increase water permeability, decrease phase transition temperature, reduce enthalpy of transition, and induce packing disorder in the DOPC membrane with an increasing concentration. On the contrary, daidzein decreases water permeability and shows negligible impact on thermodynamic parameters and packing disorder at comparable concentrations. For a cholesterol-containing DOPC bilayer, both genistein and daidzein exhibit an overall less pronounced effect on transbilayer water permeability. Their respective differential abilities to modify the physical and structural properties of biomembranes with varying lipid compositions signify a complex and sensitive nature to isoflavone interactions, which depends on the initial state of bilayer packing and the differences in the molecular structures of these soy isoflavones, and provide insights in understanding the interactions of these molecules with cellular membranes.

Understanding the Mechanical Properties of Ultradeformable Liposomes Using Molecular Dynamics Simulations.

Improving drug delivery efficiency to solid tumor sites is a central challenge in anticancer therapeutic research. Our previous experimental study (Guo et al., Nat. Commun. 2018, 9, 130) showed that soft, elastic liposomes had increased uptake and accumulation in cancer cells and tumors in vitro and in vivo respectively, relative to rigid particles. As a first step toward understanding how liposomes' molecular structure and composition modulates their elasticity, we performed all-atom and coarse-grained classical molecular dynamics (MD) simulations of lipid bilayers formed by mixing a long-tailed unsaturated phospholipid with a short-tailed saturated lipid with the same headgroup. The former types of phospholipids considered were 1,2-dioleoyl-sn-glycero-3-phosphocholine (DOPC) and 1,2-dipalmitoleoyl-sn-glycero-3-phosphocholine (termed here DPMPC). The shorter saturated lipids examined were 1,2-diheptanoyl-sn-glycero-3-phosphocholine (DHPC), 1,2-didecanoyl-sn-glycero-3-phosphocholine (DDPC), 1,2-dilauroyl-sn-glycero-3-phosphocholine (DLPC), and 1,2-dimyristoyl-sn-glycero-3-phosphocholine (DMPC). Several lipid concentrations and surface tensions were considered. Our results show that DOPC or DPMPC systems having 25-35 mol % of the shortest lipids DHPC or DDPC are the least rigid, having area compressibility moduli KA that are ∼10% smaller than the values observed in pure DOPC or DPMPC bilayers. These results agree with experimental measurements of the stretching modulus and lysis tension in liposomes with the same compositions. These mixed systems also have lower areas per lipid and form more uneven x-y interfaces with water, the tails of both primary and secondary lipids are more disordered, and the terminal methyl groups in the tails of the long lipid DOPC or DPMPC wriggle more in the vertical direction, compared to pure DOPC or DPMPC bilayers or their mixtures with the longer saturated lipid DLPC or DMPC. These observations confirm our hypothesis that adding increasing concentrations of the short unsaturated lipid DHPC or DDPC to DOPC or DPMPC bilayers alters lipid packing and thus makes the resulting liposomes more elastic and less rigid. No formation of lipid nanodomains was noted in our simulations, and no clear trends were observed in the lateral diffusivities of the lipids as the concentration, type of secondary lipid, and surface tension were varied.

Micro- and Nanoplastics Breach the Blood-Brain Barrier (BBB): Biomolecular Corona's Role Revealed.

Humans are continuously exposed to polymeric materials such as in textiles, car tires and packaging. Unfortunately, their break down products pollute our environment, leading to widespread contamination with micro- and nanoplastics (MNPs). The blood-brain barrier (BBB) is an important biological barrier that protects the brain from harmful substances. In our study we performed short term uptake studies in mice with orally administered polystyrene micro-/nanoparticles (9.55 µm, 1.14 µm, 0.293 µm). We show that nanometer sized particles-but not bigger particles-reach the brain within only 2 h after gavage. To understand the transport mechanism, we performed coarse-grained molecular dynamics simulations on the interaction of DOPC bilayers with a polystyrene nanoparticle in the presence and absence of various coronae. We found that the composition of the biomolecular corona surrounding the plastic particles was critical for passage through the BBB. Cholesterol molecules enhanced the uptake of these contaminants into the membrane of the BBB, whereas the protein model inhibited it. These opposing effects could explain the passive transport of the particles into the brain.

Entrapment and Voltage-Driven Reorganization of Hydrophobic Nanoparticles in Planar Phospholipid Bilayers.

Engineered nanoparticles (NPs) possess diverse physical and chemical properties, which make them attractive agents for targeted cellular interactions within the human body. Once affiliated with the plasma membrane, NPs can become embedded within its hydrophobic core, which can limit the intended therapeutic functionality and affect the associated toxicity. As such, understanding the physical effects of embedded NPs on a plasma membrane is critical to understanding their design and clinical use. Here, we demonstrate that functionalized, hydrophobic gold NPs dissolved in oil can be directly trapped within the hydrophobic interior of a phospholipid membrane assembled using the droplet interface bilayer technique. This approach to model membrane formation preserves lateral lipid diffusion found in cell membranes and permits simultaneous imaging and electrophysiology to study the effects of embedded NPs on the electromechanical properties of the bilayer. We show that trapped NPs enhance ion conductance and lateral membrane tension in 1,2-dioleoyl-sn-glycero-3-phosphocholine (DOPC) and 1,2-diphytanoyl-sn-glycero-3-phosphocholine (DPhPC) bilayers while lowering the adhesive energy of the joined droplets. Embedded NPs also cause changes in bilayer capacitance and area in response to applied voltage, which are nonmonotonic for DOPC bilayers. This electrophysical characterization can reveal NP entrapment without relying on changes in membrane thickness. By evaluating the energetic components of membrane tension under an applied potential, we demonstrate that these nonmonotonic, voltage-dependent responses are caused by reversible clustering of NPs within the unsaturated DOPC membrane core; aggregates form spontaneously at low voltages and are dispersed by higher transmembrane potentials of magnitude similar to those found in the cellular environment. These findings allow for a better understanding of lipid-dependent NP interactions, while providing a platform to study relationships between other hydrophobic nanomaterials and organic membranes.

Driven Engulfment of Janus Particles by Giant Vesicles in and out of Thermal Equilibrium.

The interaction between Janus colloids and giant lipid vesicles was experimentally investigated to elucidate the dynamics and mechanisms related to microparticle engulfment by lipid vesicles. Janus (Pt–SiO2 and Pt–MF, where MF is melamine formaldehyde) colloids do not spontaneously adhere to POPC or DOPC bilayers, but by applying external forces via centrifugation we were able to force the contact between the particles and the membranes, which may result in a partial engulfment state of the particle. Surface properties of the Janus colloids play a crucial role in the driven particle engulfment by vesicles. Engulfment of the silica and platinum regions of the Janus particles can be observed, whereas the polymer (MF) region does not show any affinity towards the lipid bilayer. By using fluorescence microscopy, we were able to monitor the particle orientation and measure the rotational dynamics of a single Janus particle engulfed by a vesicle. By adding hydrogen peroxide to the solution, particle self-propulsion was used to perform an active transport of a giant vesicle by a single active particle. Finally, we observe that partially engulfed particles experience a membrane curvature-induced force, which pushes the colloids towards the bottom where the membrane curvature is the lowest.

Regulation of Gramicidin Channel Function Solely by Changes in Lipid Intrinsic Curvature.

Membrane protein function is regulated by the lipid bilayer composition. In many cases the changes in function correlate with changes in the lipid intrinsic curvature (c0), and c0 is considered a determinant of protein function. Yet, water-soluble amphiphiles that cause either negative or positive changes in curvature have similar effects on membrane protein function, showing that changes in lipid bilayer properties other than c0 are important—and may be dominant. To further investigate the mechanisms underlying the bilayer regulation of protein function, we examined how maneuvers that alter phospholipid head groups effective “size”—and thereby c0—alter gramicidin (gA) channel function. Using dioleoylphospholipids and planar bilayers, we varied the head groups’ physical volume and the electrostatic repulsion among head groups (and thus their effective size). When 1,2-dioleyol-sn-glycero-3-phosphocholine (DOPC), was replaced by 1,2-dioleyol-sn-glycero-3-phosphoethanolamine (DOPE) with a smaller head group (causing a more negative c0), the channel lifetime (τ) is decreased. When the pH of the solution bathing a 1,2-dioleyol-sn-glycero-3-phosphoserine (DOPS) bilayer is decreased from 7 to 3 (causing decreased head group repulsion and a more negative c0), τ is decreased. When some DOPS head groups are replaced by zwitterionic head groups, τ is similarly decreased. These effects do not depend on the sign of the change in surface charge. In DOPE:DOPC (3:1) bilayers, pH changes from 5→9 to 5→0 (both increasing head group electrostatic repulsion, thereby causing a less negative c0) both increase τ. Nor do the effects depend on the use of planar, hydrocarbon-containing bilayers, as similar changes were observed in hydrocarbon-free lipid vesicles. Altering the interactions among phospholipid head groups may alter also other bilayer properties such as thickness or elastic moduli. Such changes could be excluded using capacitance measurements and single channel measurements on gA channels of different lengths. We conclude that changes gA channel function caused by changes in head group effective size can be predicted from the expected changes in c0.

Interactions of Bacterial Quorum Sensing Signals with Model Lipid Membranes: Influence of Acyl Tail Structure on Multiscale Response.

Many common bacteria use amphiphilic N-acyl-L-homoserine lactones (AHLs) as signaling molecules to coordinate group behaviors at high cell densities. Past studies demonstrate that AHLs can adsorb to and promote the remodeling of lipid membranes in ways that could underpin cell-cell or host-cell interactions. Here, we report that changes in AHL acyl tail group length and oxidation state (e.g., the presence or absence of a 3-oxo group) can lead to differences in the interactions of eight naturally occurring AHLs in solution and in contact with model lipid membranes. Our results reveal that the presence of a 3-oxo group impacts remodeling when AHLs are placed in contact with supported lipid bilayers (SLBs) of the phospholipid 1,2-dioleoyl-sn-glycero-3-phosphocholine (DOPC). Whereas AHLs that have 3-oxo groups generally promote the formation of microtubules, AHLs that lack 3-oxo groups generally form hemispherical caps on the surfaces of SLBs. These results are interpreted in terms of the time scales on which AHLs translocate across bilayers to relieve asymmetrical bilayer stress. Quartz crystal microbalance with dissipation measurements also reveal that 3-oxo AHLs associate with DOPC bilayers to a greater extent than their non-3-oxo analogues. In contrast, we observed no monotonic relationship between AHL tail length and bilayer reformation. Finally, we observed that 3-oxo AHLs facilitate greater transport or leakage of molecular cargo across the membranes of DOPC vesicles relative to AHLs without 3-oxo groups, also suggesting increased bilayer disruption and destabilization. These fundamental studies hint at interactions and associated multiscale phenomena that may inform current interpretations of the behaviors of AHLs in biological contexts. These results could also provide guidance useful for the design of new classes of synthetic materials (e.g., sensor elements or drug delivery vehicles) that interact with or respond selectively to communities of bacteria that use 3-oxo AHLs for cell-cell communication.

Impact of macromolecular crowding on the mesomorphic behavior of lipid self-assemblies

Using LAURDAN fluorescence we observed that water dynamics measured at the interface of DOPC bilayers can be differentially regulated by the presence of crowded suspensions of different proteins (HSA, IgG, Gelatin) and PEG, under conditions where the polymers are not in direct molecular contact with the lipid interface. Specifically, we found that the decrease in water dipolar relaxation at the membrane interface correlates with an increased fraction of randomly oriented (or random coil) configurations in the polymers, as Gelatin > PEG > IgG > HSA. By using the same experimental strategy, we also demonstrated that structural transitions from globular to extended conformations in proteins can induce transitions between lamellar and non-lamellar phases in mixtures of DOPC and monoolein. Independent experiments using Raman spectroscopy showed that aqueous suspensions of polymers exhibiting high proportions of randomly oriented conformations display increased fractions of tetracoordinated water, a configuration that is dominant in ice. This indicates a greater capacity of this type of structure for polarizing water and consequently reducing its chemical activity. This effect is in line with one of the tenets of the Association Induction Hypothesis, which predicts a long-range dynamic structuring of water molecules via their interactions with proteins (or other polymers) showing extended conformations. Overall, our results suggest a crucial role of water in promoting couplings between structural changes in macromolecules and supramolecular arrangements of lipids. This mechanism may be of relevance to cell structure/function when the crowded nature of the intracellular milieu is considered.

Illuminating Disorder Induced by Glu in a Stable Arg-Anchored Transmembrane Helix.

Membrane proteins are vital for biological function and are complex to study. Even in model peptide-lipid systems, the combined influence or interaction of pairs of chemical groups still is not well understood. Disordered proteins, whether in solution or near lipid membranes, are an emerging paradigm for the initiation and control of biological function. The disorder can involve molecular orientation as well as molecular folding. This paper reports an astonishing induction of disorder when one Glu residue is introduced into a highly stable 23-residue transmembrane helix. The parent helix is anchored by a single Arg residue, tilted at a well-defined angle with respect to the DOPC bilayer normal and undergoes rapid cone precession. When Glu is introduced two residues away from Arg, with 200° (or 160°) radial separation, the helix properties change radically to exhibit a multiplicity of three or more disordered states. The helix characteristics have been monitored by deuterium (2H) NMR spectroscopy as functions of the pH and lipid bilayer composition. The disordered multistate behavior of the (Glu, Arg)-containing helix varies with the lipid bilayer thickness and pH. The results highlight a fundamental induction of protein multistate properties by a single Glu residue in a lipid membrane environment.

Characterization of cisplatin/membrane interactions by QM/MM energy decomposition analysis

The interactions that control the permeation of cisplatin through a DOPC bilayer are unveiled by a QM/MM EDA scheme.

Self-consistent field modeling of mesomorphic phase changes of monoolein and phospholipids in response to additives.

Mapping the topological phase behaviour of lipids in aqueous solution is time consuming and finding the ideal lipid system for a desired application is often a matter of trial and error. Modelling techniques that can accurately predict the mesomorphic phase behaviour of lipid systems are therefore of paramount importance. Here, the self-consistent field theory of Scheutjens and Fleer (SF-SCF) in which a lattice refinement has been implemented, is used to scrutinize how various additives modify the self-assembled phase behaviour of monoolein (MO) and 1,2-dioleoyl-phosphatidylcholine (DOPC) lipids in water. The mesomorphic behaviour is inferred from trends in the mechanical properties of equilibrium lipid bilayers with increasing additive content. More specifically, we focus on the Helfrich parameters, that is, the mean and Gaussian bending rigidities (κ and [small kappa, Greek, macron], respectively) supplemented with the spontaneous curvature of the monolayer (Jm0). We use previously established interaction parameters that position the unperturbed DOPC system in the lamellar Lα phase ([small kappa, Greek, macron] < 0, κ > 0 and Jm0 ≈ 0). Similar interaction parameters position the MO system firmly in a bicontinuous cubic phase ([small kappa, Greek, macron] > 0). In line with experimental data, a mixture of MO and DOPC tends to be in one of these two phases, depending on the mixing ratio. Moreover we find good correlations between predicted trends and experimental data concerning the phase changes of MO in response to a wide range of additives. These correlations give credibility to the use of SF-SCF modelling as a valuable tool to quickly explore the mesomorphic phase space of (phospho)lipid bilayer systems including additives.

Cholesterol Effect on the Specific Capacitance of Submicrometric DOPC Bilayer Patches Measured by in-Liquid Scanning Dielectric Microscopy.

The specific capacitance of biological membranes is a key physical parameter in bioelectricity that also provides valuable physicochemical information on composition, phase, or hydration properties. Cholesterol is known to modulate the physicochemical properties of biomembranes, but its effect on the specific capacitance has not been fully established yet. Here we use the high spatial resolution capabilities of in-liquid scanning dielectric microscopy in force detection mode to directly demonstrate that DOPC bilayer patches at 50% cholesterol concentration show a strong reduction of their specific capacitance with respect to pure DOPC bilayer patches. The reduction observed (∼35%) cannot be explained by the small increase in bilayer thickness (∼16%). We suggest that the reduction of the specific capacitance might be due to the dehydration of the polar head groups caused by the insertion of cholesterol molecules in the bilayer. The results reported confirm the potential of in-liquid SDM to study the electrical and physicochemical properties of lipid bilayers at very small scales (down to ∼200 nm here), with implications in fields such as biophysics, bioelectricity, biochemistry, and biosensing.

Unassisted N-acetyl-phenylalanine-amide transport across membrane with varying lipid size and composition: kinetic measurements and atomistic molecular dynamics simulation

Biological membranes are essential to preserve structural integrity and regulate functional properties through the permeability of nutrients, pharmaceutical drugs, and neurotransmitters of a living cell. The movement of acetylated and amidated phenylalanine (NAFA) across 1,2-dioleoyl-sn-glycero-3-phosphocholine (DOPC) and 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine (POPC) membrane bilayers is investigated to probe physical transport. The rate of transport is measured experimentally applying parallel artificial membrane permeation assay (PAMPA). At the physiological temperature, 310 K, the measured time constants in the neutral pH were ∼6 h in DOPC and ∼3 h in POPC, while in a more acidic condition, at a pH 4.8, the time constants were ∼8 h in both lipids. Computationally, we have expanded our transport study of three aromatic dipeptides across a bilayer composed of DOPC18. In this study, we have examined the effects of lipid composition and bilayer size on the passive transport of NAFA by simulating the dipeptide in three different bilayers, with 50 DOPC lipids, 50 POPC lipids, and 40 POPC molecules. Specifically, atomistic molecular dynamics simulations with umbrella sampling were used to calculate the potential of mean force for the passive permeation of NAFA across the bilayers. Diffusion constants were then calculated by numerically solving the Smoluchowski equation. Permeability coefficients and mean first passage times were then calculated. Structural properties – Ramachandran plots, sidechain torsions, peptide insertion angles, radial distribution functions, and proximal peptide water molecules – were also examined to determine the effect of system size and lipid type. In terms of systems size, we observed a small decrease in the highest barrier of the potential of mean force and fewer sampled sidechain dihedral angle conformations with 40 versus 50 POPC lipids due to weaker membrane deformations within a smaller lipid bilayer. In terms of lipid type, DOPC contains two monounsaturated acyl chains compared to only one such acyl chain in POPC; therefore, DOPC bilayers are less ordered and more easily deformed, as seen by a much broader potential of mean force profile. The NAFA in DOPC lipid also transitioned to an internally hydrogen-bonded backbone conformation at lower membrane depths than in POPC. Similarly, as for other aromatic dipeptides, NAFA tends to insert into the membrane sidechain-first, remains mostly desolvated in the membrane center, and exhibits slow reorientations within the bilayer in both DOPC and POPC. With a joint experimental and computational study we have gained a new insight into the rate of transport and the underlying microscopic mechanism in different lipid bilayer conditions of the simplest hydrophobic aromatic dipeptide. Communicated by Ramaswamy H. Sarma

Effect of cholesterol on the fluidity of supported lipid bilayers

The lateral mobility serves as a key function of the cell membrane and can be regulated by the cholesterol. Here supported lipid bilayers were used to quantitatively analyse the influence of cholesterol on the fluidity of lipid bilayer in the environments with and without an electric field. We observed that with the increase of cholesterol proportion (0−30 mol%), the diffusion coefficient of DOPC lipid bilayer gradually decreased from 1.30 ± 0.15 μm2 s−1 to 0.28 ± 0.13 μm2 s−1, where as that of DPPC lipid bilayer increased slightly from ∼0 to 0.45 ± 0.18 μm2 s−1. We then showed that cholesterol also regulated the movement of charged lipids in the bilayer in the electric field. The migration rate of charged lipids in the DOPC bilayers slowed down as the cholesterol increased. The clarification of the dose-effect relationship of cholesterol for the bidirectional regulating effect on the mobility of lipid bilayer will extend applications of supported lipid bilayers as the model of cell membrane.

The mechanisms of action of water-soluble aminohexanoic and malonic adducts of fullerene C60 with hexamethonium on model lipid membranes

In an attempt to understand the possibility of applications of the fullerene-based systems for transporting various polar compounds like hexamethonium through the blood–brain barrier, we studied the influence of a series of derivatives of fullerene C60 in the form of salts with hexamethonium bis-anion, namely the adducts of fullerenols with 6-aminohexanoic acid (IEM-2197), and two bis-adduct malonic acid derivatives of fullerene with addents bound in two hemispheres (IEM-2143) and in equatorial positions (IEM-2144), on model membranes. We showed that IEM-2197 induced the disintegration of the bilayers composed of DOPC at the concentrations more than 2 mg/ml. IEM-2144 and IEM-2143-induced ion-permeable pores at concentrations of 0.3 and 0.02 mg/ml, respectively; herewith, IEM-2143 was characterized by the greater efficiency than IEM-2144. IEM-2197 did not significantly affect the phase behavior of DPPC, while the melting temperature significantly decreased with addition of IEM-2144 and IEM-2143. The increase in the half-width of the main transition peaks by more than 2.0 °C in the presence of IEM-2144 and IEM-2143 was observed, along with the pronounced peak deconvolution. We proposed that the immersion of IEM-2144 and IEM-2143 into the polar region of the DOPC or DPPC bilayers led to an increase in the relative mobility of tails and formation of ion-permeable defects. IEM-2197 demonstrated the more pronounced effects on the melting and ion permeability of PG- and PS-containing bilayers compared to PC-enriched membranes. These results indicated that IEM-2197 preferentially interacts with the negatively charged lipids compared to neutral species.

Partition and Solubilization of Phospholipid Vesicles by Noncovalently Constructed Oligomeric-like Surfactants.

This work has investigated the interaction of 1,2-dioleoyl-sn-glycero-3-phosphocholine (DOPC) vesicles with oligomeric surfactants noncovalently formed by sodium dodecyl sulfate (SDS) and a series of polyamines, 1,3-diaminopropane (PDA), triamine, spermidine, and spermine. The partition coefficients (P) of these surfactants between lipid bilayers and the aqueous phase are measured by isothermal titration microcalorimetry (ITC), showing that the P value increases and the Gibbs free energy of the partition becomes more negative with increasing oligomerization degree of the surfactants. This changing trend is similar to that of synthetic oligomeric surfactants regardless of the charge properties, suggesting that the polyamine and SDS molecules interact with the DOPC bilayer simultaneously. Meanwhile, the DOPC solubilization by these surfactants is evaluated by the effective surfactant-to-lipid molar ratios for the onset (Resat) and end (Resol) of the solubilization process, which are determined from the phase boundaries obtained by ITC, turbidity, and dynamic light scattering measurements. With the increment of oligomerization degree, the Resat and Resol values increase anomalously and are much larger than those of the synthetic surfactants with the same oligomerization degree, suggesting that noncovalently constructed oligomeric surfactants exhibit lower solubilization ability to phospholipid vesicles than the corresponding covalent oligomeric surfactants. Therefore, the noncovalently constructed oligomeric-like surfactants facilitate strong partition but weak solubilization to phospholipid vesicles, which may provide a useful strategy to mildly adjust the permeation and fluidity of phospholipid vesicles with solubilization delay.

Binding of the Anti-FIV Peptide C8 to Differently Charged Membrane Models: From First Docking to Membrane Tubulation.

Gp36 is the virus envelope glycoproteins catalyzing the fusion of the feline immunodeficiency virus with the host cells. The peptide C8 is a tryptophan-rich peptide corresponding to the fragment 770W-I777 of gp36 exerting antiviral activity by binding the membrane cell and inhibiting the virus entry. Several factors, including the membrane surface charge, regulate the binding of C8 to the lipid membrane. Based on the evidence that imperceptible variation of membrane charge may induce a dramatic effect in several critical biological events, in the present work we investigate the effect induced by systematic variation of charge in phospholipid bilayers on the aptitude of C8 to interact with lipid membranes, the tendency of C8 to assume specific conformational states and the re-organization of the lipid bilayer upon the interaction with C8. Accordingly, employing a bottom-up multiscale protocol, including CD, NMR, ESR spectroscopy, atomistic molecular dynamics simulations, and confocal microscopy, we studied C8 in six membrane models composed of different ratios of zwitterionic/negatively charged phospholipids. Our data show that charge content modulates C8-membrane binding with significant effects on the peptide conformations. C8 in micelle solution or in SUV formed by DPC or DOPC zwitterionic phospholipids assumes regular β-turn structures that are progressively destabilized as the concentration of negatively charged SDS or DOPG phospholipids exceed 40%. Interaction of C8 with zwitterionic membrane surface is mediated by Trp1 and Trp4 that are deepened in the membrane, forming H-bonds and cation-π interactions with the DOPC polar heads. Additional stabilizing salt bridge interactions involve Glu2 and Asp3. MD and ESR data show that the C8-membrane affinity increases as the concentration of zwitterionic phospholipid increases. In the lipid membrane characterized by an excess of zwitterionic phospholipids, C8 is adsorbed at the membrane interface, inducing a stiffening of the outer region of the DOPC bilayer. However, the bound of C8 significantly perturbs the whole organization of lipid bilayer resulting in membrane remodeling. These events, measurable as a variation of the bilayer thickness, are the onset mechanism of the membrane fusion and vesicle tubulation observed in confocal microscopy by imaging zwitterionic MLVs in the presence of C8 peptide.

Molecular Dynamics Simulations of the Elusive Matrix‐Open State of Mitochondrial ADP/ATP Carrier

AbstractWe studied and compared in detail an elusive matrix open (m‐state) and cytoplasmic open (c‐state) state of ADP/ATP carrier (AAC) protein embedded in the DOPC bilayer by microsecond molecular dynamics (MD) simulations. We analyzed both states with and without cardiolipin (CDL) molecules, with a special emphasis on the recently obtained crystallographic structure of the AAC m‐state. The obtained results show that both states of the protein are stable in the DOPC bilayer and impermeable to water. In comparison with the c‐state of AAC, the m‐state is more dynamic, but at the same time possesses a larger occluded area in the protein cavity. Both states of the protein are less flexible in simulations when CDL molecules are present, which is especially visible for the m‐state. Finally, the analysis of the protein conformational changes during MD simulations shows that protein parts at the protein/lipid boundaries are prone to larger conformational changes in contrast to central region of the protein embedded in the bilayer core, thus further supporting the cycling mechanism suggested for ADP/ATP exchange by AAC.