Properties Of Lipid Membranes Research Articles

Effect of citral partitioning on structural and mechanical properties of lipid membranes

Effect of citral partitioning on structural and mechanical properties of lipid membranes

Lipidome Unsaturation Affects the Morphology and Proteome of the Drosophila Eye.

Organisms respond to dietary and environmental challenges by altering the molecular composition of their glycerolipids and glycerophospholipids (GPLs), which may favorably adjust the physicochemical properties of lipid membranes. However, how lipidome changes affect the membrane proteome and, eventually, the physiology of specific organs is an open question. We addressed this issue in Drosophila melanogaster, which is not able to synthesize sterols and polyunsaturated fatty acids but can acquire them from food. We developed a series of semisynthetic foods to manipulate the length and unsaturation of fatty acid moieties in GPLs and singled out proteins whose abundance is specifically affected by membrane lipid unsaturation in the Drosophila eye. Unexpectedly, we identified a group of proteins that have muscle-related functions and increased their abundances under unsaturated eye lipidome conditions. In contrast, the abundance of two stress response proteins, Turandot A and Smg5, is decreased by lipid unsaturation. Our findings could guide the genetic dissection of homeostatic mechanisms that maintain visual function when the eye is exposed to environmental and dietary challenges.

Exploring the Physical Properties of Lipid Membranes with Polyhydroxy Oxanorbornane Head Group Using NBD-Conjugated and DPH Fluorescent Probes.

The present study focuses on exploring the physical properties of lipid membranes based on the polyhydroxy oxanorbornane (PH-ONB) headgroup, designed as synthetic analogues of naturally occurring archaeal lipid membranes. Specifically, we study two variants of PH-ONB headgroup-based lipids differing in the number of hydroxy groups present in the headgroup, with one having two hydroxy groups (ONB-2OH) and the other having three (ONB-3OH). These lipids form stable bilayer membranes. The study begins with a comprehensive analysis of the fluorescence characteristics of nitrobenzoxadiazole (NBD)-tagged ONB-based lipids in different solvent environments and within a model lipid membrane 1,2-dimyristoyl-sn-glycero-3-phosphocholine (DMPC). Subsequently, the physical properties of the ONB-based membranes were examined by using an NBD-tagged ONB-based probe and a commonly used extrinsic 1,6-diphenyl-1,3,5-hexatriene (DPH) fluorescent probe. The steady-state and time-resolved fluorescence properties of the NBD-tagged ONB-based probe and DPH were used to compare the physical properties of the ONB-based membranes, including polarity, fluidity, phase transition, order, hydration, location, heterogeneity, and rotational diffusion. The solid gel to liquid crystalline phase transition temperatures of ONB-2OH and ONB-3OH lipid membranes are found to be (68 ± 1) °C and (74 ± 1) °C, respectively. The variation in organization (size), fluidity, and phase transition temperature of ONB-based lipid membranes is explained by the extent of hydrogen bonding interactions between lipid head groups. ONB-based membranes exhibit characteristics similar to those of phospholipid membranes and possess a notably high phase transition temperature. These properties make them a promising and cost-effective synthetic alternative to archaeal lipid membranes with a wide range of potential applications.

Interaction of KLAKLAK-NH2 and Analogs with Biomimetic Membrane Models.

Specifically designed peptide mimetics offer higher selectivity regarding their toxicity to mammalian cells. In addition to the α-helix conformation, the specific activity is related to the peptide's ability to penetrate the cell membrane. The alterations in lipid membrane properties were addressed in the presence of the peptide KLAKLAK-NH2 and analogs containing β-alanine, strengthening the antibacterial activity and/or naphtalimide with proven anticancer properties. The molecular interactions of the peptide mimetics with POPC bilayers were studied using FTIR-ATR spectroscopy. The thermal shape fluctuation analysis of quasispherical unilamellar vesicles was applied to probe the membrane bending elasticity. The impedance characteristics of bilayer lipid membranes were measured using fast Fourier-transform electrochemical impedance spectroscopy. A lateral peptide association with the membrane is reported for β-alanine-containing peptides. The most pronounced membrane softening is found for the NphtG-KLβAKLβAK-NH2 analog containing both active groups that corroborate with the indications for 1,8-naphthalimide penetration in the lipid hydrophobic area obtained from the FTIR-ATR spectra analysis. The β-alanine substitution induces strong membrane-rigidifying properties even at very low concentrations of both β-alanine-containing peptides. The reported results are expected to advance the progress in tailoring the pharmacokinetic properties of antimicrobial peptides with strengthened stability towards enzymatic degradation. The investigation of the nonspecific interactions of peptides with model lipid membranes is featured as a useful tool to assess the antitumor and antimicrobial potential of new peptide mimetics.

The presence of free palmitic acid modulates the effects of lutein on structural and dynamic properties of lipid membranes

Free fatty acids, like palmitic acid (PA), and xanthophyll pigments, like lutein (LUT) are the natural membrane compounds in plants. To study the effect of PA on LUT and their organization, a model membrane of 1,2-dimyristoyl-sn-glycerol-3-phosphocholine (DMPC) enriched with 2 mol% PA and 1 mol% LUT was formed. Molecular mechanisms underlying the interaction between these two compounds were examined with application of molecular spectroscopy techniques, e.g., visible spectroscopy, electron paramagnetic resonance and Fourier transform infrared. We determined the monomeric/dimeric organization of LUT in the membrane. We proved that the presence of PA in the lipid phase facilitated and stabilized the formation of LUT structures in the membrane. Lutein with PA did not form strong molecular aggregates like H- and J-structures. We presented the simplified model membrane that could be a suitable representation of the physiological process of de-esterification of PA from LUT appearing in natural biomembranes in humans.

Physical insights into biological memory using phospholipid membranes.

Electrical signals may propagate along neuronal membranes in the brain, thus enabling communication between nerve cells. In doing so, lipid bilayers, fundamental scaffolds of all cell membranes, deform and restructure in response to such electrical activity. These changes impact the electromechanical properties of the membrane, which then physically store biological memory. This memory can exist either over a short or long period of time. Traditionally, biological memory is defined by the strengthening or weakening of transmissions between individual neurons. Here, we show that electrical stimulation may also alter the properties of the lipid membrane, thus pointing toward a novel mechanism for memory storage. Furthermore, based on the analysis of existing electrophysiological data, we study molecular mechanisms underlying the long-term potentiation in phospholipid membranes. Finally, we examine possible relationships between the memory capacitive properties of lipid membranes, neuronal learning, and memory.

The intricate link between membrane lipid structure and composition and membrane structural properties in bacterial membranes.

It is now evident that the cell manipulates lipid composition to regulate different processes such as membrane protein insertion, assembly and function. Moreover, changes in membrane structure and properties, lipid homeostasis during growth and differentiation with associated changes in cell size and shape, and responses to external stress have been related to drug resistance across mammalian species and a range of microorganisms. While it is well known that the biomembrane is a fluid self-assembled nanostructure, the link between the lipid components and the structural properties of the lipid bilayer are not well understood. This perspective aims to address this topic with a view to a more detailed understanding of the factors that regulate bilayer structure and flexibility. We describe a selection of recent studies that address the dynamic nature of bacterial lipid diversity and membrane properties in response to stress conditions. This emerging area has important implications for a broad range of cellular processes and may open new avenues of drug design for selective cell targeting.

Aβ Amyloid Fibers Drastically Alter the Topography and Mechanical Properties of Lipid Membranes.

Alzheimer's disease (AD) is related to the fibrillation of the Aβ peptides at neuronal membranes, a process that depends on the lipid composition and may impart different physical states to the membrane. In the present work, we study the properties of the Aβ peptide when mixed with a zwitterionic lipid (DMPC), using the Langmuir monolayer technique as an approach to control membrane physical conditions. First, we build on previous characterizations of pure Aβ monolayers and observe that, in addition to high shear, these films present a pronounced compressional hysteresis. When Aβ is assembled with DMPC in a binary film, the resulting membranes become heterogeneous, with a peptide-enriched phase distributed in a network-like pattern, and they exhibit a lateral transition that depends on the Aβ content. At lower peptide proportions, the films segregate into two well-defined phases: one consisting of lipids and another enriched with peptides. The reflectivity of these phases differs from that obtained for pure Aβ films. Thus, the formed fibers effectively cover most of the interface area and remain stable at higher pressures (from 20 to 30 mN m-1 depending on Aβ content) compared to pure peptide films (17 mN m-1). Furthermore, such structures induce a compressional hysteresis in the film, similar to that of pure peptide films (which is nonexistent in the pure lipid monolayer), even at low peptide proportions. We claim that the mechanical properties at the interface are governed by the size of the fibril-like structures. Based on the low molar fractions and surface packing at which these phenomena were observed, we postulate that as a consequence of peptide intermolecular interactions, Aβ may have drastic effects on the molecular arrangement and mechanical properties of a lipid membrane.

Study of the correlation between the structure of selected triester of phosphatidylcholine and their impact on physicochemical properties of model mammalian membranes

Cationic lipids are synthetic compounds of amphiphilic character used in Drug Delivery Systems (DDS), especially in gene therapy, as the carriers of genetic material. As it is known, the main limitation of the application of cationic lipids in DDS is their high cytotoxicity after in vivo administration and low bioactivity. This is probably related to not fully known the relationship between the lipid structure and its activity as well as the mechanism of lipofection or drug delivery. Therefore, in this work we determined the impact of a selected group of cationic lipids - triesters of phosphatidylcholine (Et-PCs) - differing in their hydrophobic structure on model mammalian membranes. In the research, as model systems, Langmuir monolayers and liposomes were applied. It was shown that the incorporation of Et-PCs into model mammalian membranes weakens interactions between lipids, causing the increase of fluidity, disordering degree and permeability of membrane. The destabilization of the membrane in this way can facilitate the entry of drugs, carried inside cationic liposomes, into the pathological cell. Moreover, the studies prove that the structure of the hydrophobic part of cationic lipids also affects the properties of lipid membranes.

Lipid composition and properties affect protein-mediated carotenoid uptake efficiency from membranes

Carotenoids are pigments of diverse functions ranging from coloration over light-harvesting to photoprotection. Yet, the number of carotenoid-binding proteins, which mobilize these pigments in physiological media, is limited, and the mechanisms of carotenoid mobilization are still not well understood. The same applies for the determinants of carotenoid uptake from membranes into carotenoproteins, especially regarding the dependence on the chemical properties of membrane lipids. Here, we investigate xanthophyll uptake capacity and kinetics of a paradigmatic carotenoid-binding protein, the homolog of the Orange Carotenoid Protein's C-terminal domain from Anabaena sp. PCC 7120 (AnaCTDH), using liposomes formed from defined lipid species and loaded with canthaxanthin (CAN) and echinenone (ECN), respectively. Phospholipids with different chain length and degree of saturation were investigated. The composition of carotenoid-loaded liposomes directly affected the incorporation yield and storage ratio of CAN and ECN as well as the rate of carotenoid uptake by AnaCTDH. Generally, saturated PC lipids were identified as unsuitable, and a high phase transition temperature of the lipids negatively affected the carotenoid incorporation and storage yield. For efficient carotenoid transfer, the velocity increases with increasing chain length or membrane thickness. An average transfer yield of 93 % and 43 % were obtained for the formation of AnaCTDH(CAN) and AnaCTDH(ECN) holoproteins, respectively. In summary, the most suitable lipids for the formation of AnaCTDH(CAN/ECN) holoproteins by carotenoid transfer from artificial liposomes are phosphatidylcholine (18:1) and phosphatidylglycerol (14:0). Thus, these two lipids provide the best conditions for further investigation of lipid-protein interaction and the carotenoid uptake process.

Controlling plasmonic suprastructures through self-assembly of gold nanoparticles with hybrid copolymer-lipid vesicles

Hybrid lipid membranes incorporating amphiphilic copolymers have gained significant attention due to their potential applications in various fields, including drug delivery and sensing. By combining the properties of copolymers and lipid membranes, such as enhanced chemical tunability and stability, environmental responsiveness, and multidomain nature, novel membrane architectures have been proposed. In this study, we investigated the potentialities of hybrid membranes made of two distinct components: the rigid fully saturated phospholipid 1,2-dipalmitoyl-sn-glycero-3-phosphocholine (DPPC) and the soft copolymer poly(butadiene-b-ethyleneoxide) (PBD-b-PEO). The objective was to explore the interaction of citrate-coated gold nanoparticles (AuNPs) and the hybrid membrane, aiming at constructing AuNPs-hybrid vesicles suprastructures with controlled and adjustable plasmonic properties. A series of experimental techniques were employed to investigate hybrid free-standing and supported membranes. The results revealed that the incorporation of the copolymer into the lipid membrane promotes AuNPs clustering, demonstrating a distinctive aggregative phenomenon of citrate-coated AuNPs on multidomain membranes. Importantly, we show that the size and morphology of AuNPs clusters can be precisely controlled in non-homogeneous membranes, enabling the formation of hybrid suprastructures with controlled patch properties. These results highlight the potential of lipid-copolymer hybrid membranes for designing functional materials with tailored plasmonic properties, with potential applications in nanomedicine and sensing.

Mechanical tomography of an archaeal lemon-shaped virus reveals membrane-like fluidity of the capsid and liquid nucleoprotein cargo

Archaeal lemon-shaped viruses have unique helical capsids composed of highly hydrophobic protein strands which can slide past each other resulting in remarkable morphological reorganization. Here, using atomic force microscopy, we explore the biomechanical properties of the lemon-shaped virions of Sulfolobus monocaudavirus 1 (SMV1), a double-stranded DNA virus which infects hyperthermophilic (~80 °C) and acidophilic (pH ~ 2) archaea. Our results reveal that SMV1 virions are extremely soft and withstand repeated extensive deformations, reaching remarkable strains of 80% during multiple cycles of consecutive mechanical assaults, yet showing scarce traces of disruption. SMV1 virions can reversibly collapse wall-to-wall, reducing their volume by ~90%. Beyond revealing the exceptional malleability of the SMV1 protein shell, our data also suggest a fluid-like nucleoprotein cargo which can flow inside the capsid, resisting and accommodating mechanical deformations without further alteration. Our experiments suggest a packing fraction of the virus core to be as low as 11%, with the amount of the accessory proteins almost four times exceeding that of the viral genome. Our findings indicate that SMV1 protein capsid displays biomechanical properties of lipid membranes, which is not found among protein capsids of other viruses. The remarkable malleability and fluidity of the SMV1 virions are likely necessary for the structural transformations during the infection and adaptation to extreme environmental conditions.

Exploring Fullerenol-C60(OH)24 interactions with lipid bilayers: Molecular dynamics study of agglomeration and surface deposition

In this work, the physical–chemical properties of lipid membranes interacting with Fullerenol-C60(OH)24 molecules immersed in an aqueous solution were studied using Molecular Dynamics (MD) simulations. In order to analyze the role played by the concentration of Fullerene-C60(OH)24 in the interaction with lipid membranes, we carried out simulations with variations in the number (n = 1, 2, 4, 8, 12, 16 or 20) of Fullerene-C60(OH)24 molecules present in the solution in which the lipid membrane was immersed. The lipid membrane simulated was composed of DOPC (dipalmitoylphosphatidylcholine) with 30% of cholesterol molecules (DOPC/CHOL(30%)). The average behavior of the Coulomb and vdW interaction energy shows that as the Fullerenol-C60(OH)24 molecules concentration increases, insertion of Fullerenol in the lipid bilayer becomes energetically unfavorable since they tend to interact with each other, leading to the formation of clusters. From the mass density, we confirmed that there is a decrease in the insertion of Fullerenol-C60(OH)24 molecules in the bilayer as n increases. We noticed that Fullerenol molecules get closer to each other and are deposited on the surface of the lipid bilayers but do not penetrate them. The diffusion coefficient (MSD) reveals a decrease in the mobility of Fullerenol molecules as n increases. This observation confirms the agglomeration of Fullerenol on the lipid membrane’s surface. A detailed analysis of hydrogen bonds (HB) involving Fullerenol–Fullerenol, Fullerenol–DOPC, and Fullerenol–Water pairs provides insight into this phenomenon. While the average number of HBs in the Fullerenol–Water pair remains relatively constant as n increases, there is a significant increase in HB interactions for the Fullerenol–Fullerenol pair. In contrast, the Fullerenol–DOPC pair experiences a decrease in HBs, underscoring the preference of Fullerenol to agglomerate and deposit onto the lipid membrane's surface.

The effect of trans-resveratrol on the physicochemical properties of lipid membranes with different cholesterol content

Resveratrol is one of the most popular phytoalexins, which naturally occurs in grapes and red wine. This compound not only has beneficial effects on the human body, especially on the cardiovascular system, but also has antiviral, antibacterial and antifungal properties. In addition, resveratrol may have therapeutic effects against various types of cancer. The mechanism of action of resveratrol is not fully understood, but it is suspected that one of the most important steps is its interaction with the cell membrane and changing its molecular organization. Therefore, in the present study, we investigated the effects of resveratrol at different concentrations (0–75 μM) on model membranes composed of POPC, SM and cholesterol, in systems with different cholesterol contents and a constant POPC/SM molar ratio (1:1). Our tests included systems containing 5, 15 and 33.3 mol% cholesterol. Tests were carried out for monolayers using the Langmuir monolayer technique supported by Brewster angle microscopy and penetration experiments. Bilayer (liposome) experiments included calcein release, steady-state DPH fluorescence anisotropy and partition coefficients. The results showed that resveratrol interacts with model cell membranes (lipid monolayers and lipid bilayers), and its incorporation into membranes is accompanied by changes in their physicochemical parameters, such as lipid packing, fluidity and permeability. Furthermore, we showed that the cholesterol content of the membrane significantly affects the degree of incorporation of resveratrol into the model membrane, which may indicate that the molecular mechanism of action of this compound is closely related to its interactions with lipid rafts, domains responsible for regulating various cellular functions.

The influence of Leucidal – eco-preservative from radish – on model lipid membranes and selected pathogenic bacteria

In this work the effect of Leucidal - a natural preservative from radish dedicated to be used in cosmetics - on bacteria cells and model bacteria membranes was investigated. To get insight into the mechanism of action of this formulation the lipid Langmuir monolayers imitating Escherichia coli (E. coli) and Staphylococcus aureus (S. aureus) membranes were prepared. Then, the influence of Leucidal on model systems was investigated by means of the surface pressure/area measurements, penetration studies and Brewster Angle Microscopy (BAM) visualization. Similar experiments were done also for one component monolayers formed from the model membrane lipids. The in vitro tests were done on five different bacteria species (E. coli, Enterococcus faecalis, S. aureus, Salmonella enterica, Pseudomonas aeruginosa). Leucidal was found to decrease packing of the monolayers, however, it was excluded from the films at higher concentrations. Model membrane experiments evidenced also a stronger affinity of the components of this eco-preservative to E. coli vs S. aureus membrane. Among one component films, those formed from phosphatidylglycerols and cardiolipins were more sensitive to the presence of Leucidal. However, in vitro tests evidenced that Leucidal exerts stronger inhibitory effect against S. aureus bacteria as compared to E. coli strain. These findings were discussed from the point of view of the role of Leucidal components and the lipid membrane properties in the membrane - based mechanism of action of this preservative. The results allow one to suggest that the membrane may not be the main site of action of Leucidal on bacteria. Moreover, since high concentration of the tested preparation exerted antibacterial activity in relation to all tested bacteria, a low selectivity of Leucidal can be postulated, which may be problematic from the point of view of its effect on the skin microbiome.

Biophysical Properties of Lipid Membranes from Barley Roots during Low-Temperature Exposure and Recovery.

Glycerolipid remodeling, a dynamic mechanism for plant subsistence under cold stress, has been posited to affect the biophysical properties of cell membranes. In barley roots, remodeling has been observed to take place upon exposure to chilling stress and to be partially reverted during stress relief. In this study, we explored the biophysical characteristics of membranes formed with lipids extracted from barley roots subjected to chilling stress, or during a subsequent short- or long-term recovery. Our aim was to determine to what extent barley roots were able to offset the adverse effects of temperature on their cell membranes. For this purpose, we analyzed the response of the probe Laurdan inserted in bilayers of different extracts, the zeta potential of liposomes, and the behavior of Langmuir monolayers upon compression. We found important changes in the order of water molecules, which is in agreement with the changes in the unsaturation index of lipids due to remodeling. Regarding Langmuir monolayers, we found that films from all the extracts showed a reorganization at a surface pressure that depends on temperature. This reorganization occurred with an increase in entropy for extracts from control plants and without entropy changes for extracts from acclimated plants. In summary, some membrane properties were recovered after the stress, while others were not, suggesting that the membrane biophysical properties play a role in the mechanism of plant acclimation to chilling. These findings contribute to our understanding of the impact of lipid remodeling on biophysical modifications in plant roots.

Influence of docosahexaenoic acid on the interfacial behavior of cholesterol-containing lipid membranes: Interactions with small amphiphiles and hydration properties

Cholesterol is known to significantly modify both the structural and the dynamical properties of lipid membranes. On one side, the presence of free cholesterol molecules has been determined to stiffen the membrane bilayer by stretching the hydrophobic tails. Additionally, recent experimental and computational findings have made evident the fact that cholesterol also alters the dynamics and the hydration properties of the polar head groups of DPPC model lipid membranes. In turn, we have recently shown that the Omega-3 fatty acid docosahexaenoic acid, DHA, counteracts the effect of cholesterol on DPPC membrane's mechanical properties by fluidizing the bilayer. However, such behavior represents in fact a global outcome dominated by the larger lipid hydrophobic tails that neither discriminates between the different parts of the membrane nor elucidates the effect on membrane hydration and binding properties. Thus, we now perform molecular dynamics simulations to scrutinize the influence of DHA on the interfacial behavior of cholesterol-containing lipid membranes by characterizing their hydration properties and their binding to amphiphiles. We find that while cholesterol destabilizes interactions with amphiphiles and slightly weakens the lipid's hydration layer, the incorporation of DHA practically restores the interfacial behavior of pure DPPC.

Viscosity-Sensitive Membrane Dyes as Tools To Estimate the Crystalline Structure of Lipid Bilayers.

Lipid membranes are crucial for cellular integrity and regulation, and tight control of their structural and mechanical properties is vital to ensure that they function properly. Fluorescent probes sensitive to the membrane's microenvironment are useful for investigating lipid membrane properties; however, there is currently a lack of quantitative correlation between the exact parameters of lipid organization and a readout from these dyes. Here, we investigate this relationship for "molecular rotors", or microviscosity sensors, by simultaneously measuring their fluorescence lifetime to determine the membrane viscosity, while using X-ray diffraction to determine the membrane's structural properties. Our results reveal a phase-dependent correlation between the membrane's structural parameters and mechanical properties measured by a BODIPY-based molecular rotor, giving excellent predictive power for the structural descriptors of the lipid bilayer. We also demonstrate that differences in membrane thickness between different lipid phases are not a prerequisite for the formation of lipid microdomains and that this requirement can be disrupted by the presence of line-active molecules. Our results underpin the use of membrane-sensitive dyes as reporters of the structure of lipid membranes.

Accelerating Lipid Flip-Flop at Low Concentrations: A General Mechanism for Membrane Binding Peptides.

We report a physicochemical investigation of the lipid transport properties of model lipid membranes in the presence of the antimicrobial peptide indolicidin through comparisons of experimental SANS/SAXS scattering techniques to fully atomistic molecular dynamics simulations. In agreement with the experiment, we show that upon peripheral binding of the peptides, even at low concentrations, lipid flip-flop dynamics is greatly accelerated. Computer modeling elucidates the interplay between structural changes and lipid dynamics induced by peptides and proposes a mechanism for the mode of action of antimicrobial peptides, assessing the major role of entropy for the catalysis of the flipping events. The mechanism introduced here is universal for all peptides with preferential peripheral binding to the membrane as it does not depend on the specific amino acid sequence.

Understanding the Molecular Mechanism of Anesthesia: Effect of General Anesthetics and Structurally Similar Non-Anesthetics on the Properties of Lipid Membranes.

General anesthesia can be caused by various, chemically very different molecules, while several other molecules, many of which are structurally rather similar to them, do not exhibit anesthetic effects at all. To understand the origin of this difference and shed some light on the molecular mechanism of general anesthesia, we report here molecular dynamics simulations of the neat dipalmitoylphosphatidylcholine (DPPC) membrane as well as DPPC membranes containing the anesthetics diethyl ether and chloroform and the structurally similar non-anesthetics n-pentane and carbon tetrachloride, respectively. To also account for the pressure reversal of anesthesia, these simulations are performed both at 1 bar and at 600 bar. Our results indicate that all solutes considered prefer to stay both in the middle of the membrane and close to the boundary of the hydrocarbon domain, at the vicinity of the crowded region of the polar headgroups. However, this latter preference is considerably stronger for the (weakly polar) anesthetics than for the (apolar) non-anesthetics. Anesthetics staying in this outer preferred position increase the lateral separation between the lipid molecules, giving rise to a decrease of the lateral density. The lower lateral density leads to an increased mobility of the DPPC molecules, a decreased order of their tails, an increase of the free volume around this outer preferred position, and a decrease of the lateral pressure at the hydrocarbon side of the apolar/polar interface, a change that might well be in a causal relation with the occurrence of the anesthetic effect. All these changes are clearly reverted by the increase of pressure. Furthermore, non-anesthetics occur in this outer preferred position in a considerably smaller concentration and hence either induce such changes in a much weaker form or do not induce them at all.