DOPC Monolayers Research Articles

Interactions of Alkali Ions and Phospholipid Monolayers at Organic-Water Interfaces

The self-assembled phospholipids monolayers at interfaces between two immiscible electrolyte solutions (ITIES) are commonly used to study the interactions between cell membranes and species passing in and out through membrains such as ions, drugs, and other nutrients. In this study, we utilized electrochemical techniques such as open circuit potential (OCP), cyclic voltammetry (CV), and electrochemical impedance spectroscopy (EIS) to study the pure phospholipd monolayer formed by 1,2-Dioleoyl-sn-glycero-3-phosphocholine (DOPC) and its mixture with 1,2-dioleoyl-sn-glycero-3-phospho-(1'-racglycerol) (sodium salt) (DOPG) at 1,2-dichloroethane-water interfaces. Our study has shown that at higher applied electric potentials, there seems to be no change in the facilitation of cation transfers by DOPC/DOPG monolayer compared to DOPC monolayer. However, at lower applied potentials, the DOPGs in the mixed monolayer seemed to leave the interface under the influence of the applied electric field, indicated by both the higher capacitance obtained throught the EIS measurements and the higher currents from the CV measurements compared to results obtained from the sample consisted of the pure DOPC monolayer. Furthermore, our study also showed a general rise in capacitance when incorporating heavier alkali ions in the aqueous phase for the sample with the mixture of DOPC/DOPG at higher applied electric potentials.

Interfacial Interactions of a Myoglobin/DOPC Hybrid System at the Air-Water Interface and Its Physicochemical Properties.

In the present study, the intermolecular interactions between a water-insoluble phospholipid (DOPC) and water-soluble protein (myoglobin) and the interaction among themselves were investigated at the air-water interface using the Langmuir and Langmuir-Blodgett techniques. The effects of changes in physicochemical factors, like pH and temperature, on these interactions were also examined. Surface pressure-molecular area (π-A) isotherms of the DOPC monolayer at the air-water interface, with and without myoglobin (Myo) revealed the evolution of various physical properties, such as elastic, thermodynamic, and hysteric properties, in response to changes in subphase pH and temperature. With the increment of subphase pH from 5 to 7 at a fixed temperature (20 °C), the DOPC isotherm expanded, and the in-plane elasticity (CS-1) decreased, but no significant presence of hysteresis was encountered in either of the pH values. On the other hand, a diminution of temperature (from 20 to 5 °C) leads to an expansion of monolayers yielding low elasticity and significant hysteresis. The incorporation of Myo molecules within the DOPC monolayer decreased the CS-1 value of the DOPC monolayer. Such a decrement in CS-1 was also encountered while increasing the pH and decreasing the temperature (T) of the subphase in the absence of Myo. Systematic expansion of DOPC isotherm and increased hysteric area with the increase in Myo proportion were observed and the atomic force microscopy (AFM) observations suggested a strong conjugation between Myo and DOPC in the mixed monolayer. The denaturation effect of Myo molecules was studied using AFM at different temperatures. Furthermore, the Myo molecules were found to be most surface active at pH = 7, which is very close to its isoelectric point. These observations come up with the interaction mechanism between biomolecules under dynamically varied conditions.

Interfacial interactions in urease/stearic acid and urease/DOPC binary mixed systems at the air-water and the air-solid interfaces

In the present study, the protein-lipid interactions between water-soluble metalloenzyme urease (Urs) and insoluble stearic acid (SA) and 1,2-dioleoyl-sn-glycero-3-phosphocholine (DOPC) were investigated at the air–water interface using the Langmuir-Blodgett (LB) technique. Surface pressure (π) - molecular area (A) isotherms were measured for the pure SA and pure DOPC monolayers in the presence of urease in the water subphase. From the (π – A) isotherms of the binary system, it was found that urease strongly interacts with SA and DOPC. The compressibility of the SA monolayer was reduced with the incorporation of urease; a similar trend was also found with DOPC. Moreover, an association of urease with lipid monolayers was found to be surface pressure (π) dependent. The presence of a little hump in the mixed isotherms of Urs-SA at low surface pressure supports this observation. However, the miscibility studies and the calculations of excess Gibb’s free energy indicate an attractive interaction between urease and SA molecules. The considerable morphological variations observed by atomic force microscopy (AFM) in urease-SA deposited films onto hydrophilic and hydrophobic Si(100) surfaces suggest that urease was strongly bound with the surrounding lipid SA on the hydrophilic surfaces but the urease molecules were rigorously denatured on the hydrophobic surfaces. The AFM analysis of Urs-DOPC mixed LB films at room temperature (RT) and elevated temperature suggests substantial denaturation of urease in a hydrophobic environment leading us to speculate the exposure of tryptophan moieties of the urease molecules to the hydrophobic air phase.

Studies on the interactions of tiny amounts of common ionic surfactants with unsaturated phosphocholine lipid model membranes

In order to provide the fundamental information about the interactions of common anionic surfactants with the basic unsaturated phospholipids the influence of three cationic (dodecyltrimethylammonium bromide, DTAB; tetradecyltrimethylammonium bromide, TTAB and hexadecyltrimethylamonium bromide, CTAB) and one anionic (sodium dodecylsulfate, SDS) surfactants on the properties of the 1-palmitoyl-2-oleoyl-glycero-3-phosphocholine (POPC) and 1,2-dioleoyl-sn-glycero-3-phosphocholine (DOPC) layers was investigated. The studies proved that a tiny amount of the ionic surfactant added to the already synthesized liposome suspension is sufficient to change the zeta potential of the POPC and DOPC liposomes significantly. This impact increases with the surfactant concentration, the alkyl chain length of the surfactant and the degree of lipid saturation. Moreover, this effect is greater for the anionic surfactant than for the cationic one of the same alkyl chain length. The observed findings were confirmed in the course of the research carried out with the use of the corresponding Langmuir monolayers where the surface pressure – mean area isotherms, the compressibility modulus – surface pressure dependences, the monolayer penetration tests, the surface potential – mean molecular area isotherms and Brewster angle microscopy were discussed. It was found that the presence of the surfactants shifts the isotherms towards larger molecular area, to the higher extent for the SDS than DTAB. This effect increases with the increasing surfactant concentration in the subphase. Moreover, the investigated surfactants remain in the monolayer even at high surface pressure. Nevertheless, no effect on the morphology of the POPC and DOPC monolayers was detected from the BAM images. The surface potential and surface charge of the liposomes calculated on the basis of the zeta potential results reflected the interactions between the surfactant and the lipid layers.

Bisphenol A exposure induces multiple effects in DOPC membrane models

• BPA affects the DOPC π-A isotherms without altering the lipid fluidity. • BPA surface activity has an important role in the morphology of lipid monolayers. • The interaction with lipid occurs by phosphate groups and promotes a disorder in the lipid tail. • BPA induces multiple morphology changes on giant unilamellar vesicles. • The effect of BPA on vesicles showed a size dependence. Due to its now well-known endocrine-disrupting behavior, Bisphenol A (BPA) has focused on many interactions, detection, and removal studies in the last few years. The application of simple membrane models to identify specific interactions in some regions of the cells, such as the membrane, is essential to help understand the aspects of the mechanisms of action. This work evaluated the effects of the BPA exposure over membrane models of a fluid phospholipid, the 1,2-dioleoyl- sn -glycero-3-phosphocholine (DOPC). The BPA interaction with the DOPC monolayers was performed using Langmuir films, evaluating changes in π-A isotherms, lipid fluidity, and interaction at the molecular level by PM-IRRAS spectroscopy. Giant unilamellar vesicles (GUVs) were used to reveal alteration in the shape and morphology of the lipid bilayers promoted by BPA. Multiple effects were observed in the GUVs, including surface fluctuations, an endocytosis-like effect, the formations of budding, and scissions. These alterations could be related to changes observed in the phosphate groups of DOPC in the PM-IRRAS analysis and the lipophilic properties of BPA. Furthermore, the changes observed indicate that BPA has an affinity to the membrane, which could lead to different implications or effects in cells not yet understood, which requires further investigations.

The effect of chitosan/TiO2/hyaluronic acid subphase on the behaviour of 1,2-dioleoyl-sn-glycero-3-phosphocholine membrane.

The main aim of the study was to determine the effect of two polysaccharides: chitosan (Ch) and hyaluronic acid (HA), and/or titanium dioxide (TiO2) on the structure and behaviour of the 1,2-dioleoyl-sn-glycero-3-phosphocholine (DOPC) membrane. To achieve this goal the surface pressure as a function of the area per molecule (π-A) isotherm for the phospholipid monolayer was recorded. The shape of the π-A isotherms and compression-decompression cycles, as well as the compression modulus values, were analysed in terms of biocompatibility. Besides, morphology and thickness of the phospholipid layers obtained by means of Brewster angle microscope at different π, were determined. The obtained results show that both polysaccharides Ch, HA, as well inorganic TiO2 affect slightly the structure of the DOPC monolayer but do not disrupt it. Their presence brings no typical arrangements of both the polar heads and tails of DOPC molecules at the interface.

Unsaturated lipids modulating the interaction of the antileishmanial isolinderanolide E with models of cellular membranes

The present work evaluated the antiprotozoal activity of isolinderanolide E, isolated from the Brazilian plant Nectandra oppositifolia, against promastigote forms of Leishmania (Leishmania) amazonensis. The compound exhibited an EC50 value of 20.3 μM, similar to the positive control miltefosine (IC50 of 19.4 μM), and reduced toxicity to macrophages (CC50 > 200 μM). Based on these results, Langmuir monolayers of two unsaturated lipids: 1,2-dioleoyl-sn-glycero-3-phosphocholine (DOPC) and 1,2-dioleoyl-sn-glycero-3-phosphoethanolamine (DOPE), were employed as a model of mammalian and parasite membranes, respectively, to study the interaction of isolinderanolide E at a molecular level. The films were characterized with tensiometry (surface pressure-area isotherms and surface pressure-time curves), infrared spectroscopy, and Brewster angle microscopy (BAM). This compound changed the profile of the isotherms leading to fluid DOPC and DOPE monolayers, which were not able to attain rigid states even with compression. Infrared spectroscopy showed that the bioactive compound decreases the trans/gauche ratio conformers related to the molecular conformational disorder. BAM showed the formation of specific aggregates upon drug incorporation. In conclusion, isolinderanolide E changes the thermodynamic, mechanical, structural, and morphological characteristics of the monolayer of these unsaturated lipids, which may be essential to understand the action at the molecular level bioactives in biointerfaces.

Masking of a malodorous substance on 1,2-dioleoyl-sn-glycero-3-phosphocholine molecular layer

The performance of a trans-2-nonenal (NE)-including 1,2-dioleoyl-sn-glycero-3-phosphocholine (DOPC) molecular layer as a model of epithelial cellular membrane damaged by malodorous substance was investigated to understand the masking effect of odorants (benzaldehyde (BA), cyclohexanecarboxyaldehyde (CA), 4-anisaldehyde (AA), and 4-methoxycyclohexanecarbaldehyde (MA)). The surface pressure of the DOPC monolayer increased upon the addition of NE, and the surface pressure–area isotherm of the DOPC-NE mixture was characteristically responsive to the odorants (BA, CA, AA, and MA). Differential scanning calorimetry, Fourier-transform infrared spectroscopy, and ultraviolet absorption spectrophotometry were conducted to evaluate the interactions among DOPC, NE, and the odorants. The results suggested that odorants with an aromatic ring form a complex with NE and were eliminated from the DOPC membrane. We found that odorants with an aromatic ring play an important role in inducing the masking effect on the DOPC-NE mixed monolayer because of the characteristic interactions between NE and odorant.

Interaction of Amyloid-β-(1-42) Peptide and Its Aggregates with Lipid/Water Interfaces Probed by Vibrational Sum-Frequency Generation Spectroscopy.

In this study, we use surface-sensitive vibrational sum-frequency generation (VSFG) spectroscopy to investigate the interaction between model lipid monolayers and Aβ(1-42) in its monomeric and aggregated states. Combining VSFG with atomic force microscopy (AFM) and thioflavin T (ThT) fluorescence measurements, we found that only small aggregates with probably a β-hairpin-like structure adsorbed to the zwitterionic lipid monolayer (DOPC). In contrast, larger aggregates with an extended β-sheet structure adsorbed to a negatively charged lipid monolayer (DOPG). The adsorption of small, initially formed aggregates strongly destabilized both monolayers, but only the DOPC monolayer was completely disrupted. We showed that the intensity of the amide-II' band in achiral (SSP) and chiral (SPP) polarization combinations increased in time when Aβ(1-42) aggregates accumulated at the DOPG monolayer. Nevertheless, almost no adsorption of preformed mature fibrils to DOPG monolayers was detected. By performing spectral VSFG calculations, we revealed a clear correlation between the amide-II' signal and the degree of amyloid aggregates (e.g., oligomers or (proto)fibrils) of various Aβ(1-42) structures. The calculations showed that only structures with a significant amyloid β-sheet content have a strong amide-II' intensity, in line with previous Raman studies. The combination of the presented results substantiates the amide-II(') band as a legitimate amyloid marker.

Molecular-level effects on cell membrane models to explain the phototoxicity of gold shell-isolated nanoparticles to cancer cells

Metallic nanoparticles are promising agents for photothermal cancer therapy (PTT) owing to their photostability and efficient light-to-heat conversion, but their possible aggregation remains an issue. In this paper, we report on the photoinduced heating of gold shell-isolated nanoparticles (AuSHINs) in in vitro experiments to kill human oropharyngeal (HEp-2) and breast (BT-474 and MCF-7) carcinoma cells, with cell viability reducing below 50 % with 2.2 × 1012 AuSHINs/mL and 6 h of incubation. This toxicity to cancer cells is significantly higher than in previous works with gold nanoparticles. Considering the AuSHINs dimensions we hypothesize that cell uptake is not straightforward, and the mechanism of action involves accumulation on phospholipid membranes as the PTT target for photoinduced heating and subsequent generation of reactive oxygen species (ROS). Using Langmuir monolayers as simplified membrane models, we confirmed that AuSHINs have a larger effect on 1,2-dioleoyl-sn-glycero-3-phospho-l-serine (DOPS), believed to represent cancer cell membranes, than on 1,2-dioleoyl-sn-glycero-3-phosphocholine (DOPC) taken as representative of healthy eukaryotic cells. In particular, data from polarization-modulated infrared reflection absorption spectroscopy (PM-IRRAS) revealed an increased conformational order of DOPS tails due to the stronger adsorption of AuSHINs. Furthermore, light irradiation reduced the stability of AuSHINs containing DOPC and DOPS monolayers owing to oxidative reactions triggered by ROS upon photoinduced heating. Compared to DOPC, DOPS lost nearly twice as much material to the subphase, which is consistent with a higher rate of ROS formation in the vicinity of the DOPS monolayer.

High-throughput electrochemical sensing platform for screening nanomaterial–biomembrane interactions

A high-throughput, automated screening platform has been developed for the assessment of biological membrane damage caused by nanomaterials. Membrane damage is detected using the technique of analyzing capacitance-current peak changes obtained through rapid cyclic voltammetry measurements of a phospholipid self-assembled monolayer formed on a mercury film deposited onto a microfabricated platinum electrode after the interaction of a biomembrane-active species. To significantly improve wider usability of the screening technique, a compact, high-throughput screening platform was designed, integrating the monolayer-supporting microfabricated electrode into a microfluidic flow cell, with bespoke pumps used for precise, automated control of fluid flow. Chlorpromazine, a tricyclic antidepressant, and a citrate-coated 50 nm diameter gold nanomaterial (AuNM) were screened to successfully demonstrate the platform's viability for high-throughput screening. Chlorpromazine and the AuNM showed interactions with a 1,2-dioleoyl-sn-glycero-3-phosphocholine (DOPC) monolayer at concentrations in excess of 1 µmol dm-3. Biological validity of the electrochemically measured interaction of chlorpromazine with DOPC monolayers was confirmed through quantitative comparisons with HepG2 and A549 cytotoxicity assays. The platform also demonstrated desirable performance for high-throughput screening, with membrane interactions detected in <6 min per assay. Automation contributed to this significantly by reducing the required operating skill level when using the technique and minimizing fluid consumption.

The Dependence of the Miscibility, Stability and Compressibility of L-&amp;lt;i&amp;gt;α&amp;lt;/i&amp;gt; Dioleoylphosphatidylcholine/Rutin Laurate Monolayer at the Air/Water on Temperature

The miscibility, stability and compressibility of L-α dioleoylphosphatidylcholine/rutin laurate mixed monolayer at the air/water were investigated by Langmuir film balance to reveal the characteristic of the molecular interaction. The two components of DOPC/RL mixed monolayer were miscible throughout the mixture composition range and at three experimental temperatures of 10°C, 25°C and 37°C. At all experimental conditions, RL increased the compressibility and elasticity of the DOPC monolayer. Both the temperature and the composition of the membrane affected the form of intermolecular forces in the mixed monolayer.

Role of Toluidine Blue-O Binding Mechanism for Photooxidation in Bioinspired Bacterial Membranes.

The alarming increase in bacterial resistance to antibiotics has demanded new strategies for microbial inactivation, which include photodynamic therapy whose activity relies on the photoreaction damage to the microorganism membrane. Herein, the binding mechanisms of the photosensitizer toluidine blue-O (TBO) on simplified models of bacterial membrane with Langmuir monolayers of 1,2-dioleoyl-sn-glycero-3-phosphoethanolamine (DOPE) and 1,2-dioleoyl-sn-glycero-3-phospho-(1'-rac-glycerol) (DOPG) were correlated to the effects of the photoinduced lipid oxidation. Langmuir monolayers of 1,2-dioleoyl-sn-glycero-3-phosphocholine (DOPC) were also used as a reference of mammalian membranes. The surface pressure isotherms combined with polarization-modulated infrared reflection absorption spectroscopy revealed that TBO expands DOPC, DOPE, and DOPG monolayers owing to electrostatic interactions with the negatively charged groups in the phospholipids, with a stronger adsorption on DOPG, which has a net surface charge. Light irradiation made the TBO-containing DOPC and DOPE monolayers less unstable as a result of the singlet oxygen (1O2) reaction with the chain unsaturation and hydroperoxide formation. In contrast, the decreased stability of the irradiated TBO-containing DOPG monolayer suggests the cleavage of carbon chains. The anionic nature of DOPG allowed a deeper penetration of TBO into the chain region, favoring contact-dependent reactions between the excited triplet state of TBO and lipid unsaturations or/and hydroperoxide groups, which is the key for the cleavage reactions and further membrane permeabilization.

Liposomal membrane fusion induced by extrusion method

Objective To study the application of extrusion method in inducing liposome membrane fusion or membrane component mixing, and to investigate the effect of different extrusion conditions on liposome membrane fusion rate. Methods N-(7-nitrobenz-2-oxa-1,3-diazol-4-yl) phosphatidylethanolamine (N-NBD-PE) and N-(lissamine rhodamine B sulfonyl) phosphatidylethanolamine (N-Rh-PE) labeled 1, 2-dioleoyl lecithin (DOPC) liposomes and non-fluorescently labeled DOPC monolayer liposomes were mixed and extruded. The fluorescence changes before and after the extrusion of the mixed liposomes were observed using laser scanning confocal microscope, and the membrane fusion rate of the mixed liposomes was calculated by fluorescence resonance energy transfer method. Besides, the effects of extrusion times, extrusion pressure and temperature on the fusion rate of liposome membrane were studied. Results The results of laser scanning confocal microscopy showed that the distribution density and intensity of the green fluorescence of N-NBD-PE increased significantly after the extrusion of fluorescently labeled and non-fluorescent labeled DOPC liposomes, which confirmed membrane fusion. After 75 times of extrusion treatments, the liposome membrane fusion rate can reach 26%. The number of extrusions, extrusion pressure and temperature had a significant effect on the fusion rate of the liposome membrane. The higher the number of the extrusions, the smaller the extrusion pressure and the higher the efficiency of the liposome membrane fusion were at physiological temperature. Conclusions Extrusion method can induce liposome membrane fusion and membrane component mixing, and the prepared liposome has a narrower particle size distribution, which is expected to be a new method to induce the bilayer membrane fusion of liposome or lipid vesicle. Key words: Extrusion; Liposome; Lipid bilayer fusion; Fluorescence resonance energy transfer

Evidence of photoinduced lipid hydroperoxidation in Langmuir monolayers containing Eosin Y.

Photodynamic therapy (PDT) efficiency depends on many factors including the incorporation of the photosensitizer (PS) in cell membranes and possible lipid hydroperoxidation. In this study, we show that hydroperoxidation may be photoinduced when eosin Y is incorporated into Langmuir monolayers that serve as cell membrane models. This occurs for Langmuir monolayers of 1,2-dioleoyl-sn-glycero-3-phosphocholine (DOPC) and 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine (POPC), which have unsaturation in their hydrophobic chains. In contrast, light irradiation had no effect on monolayers of saturated 1,2-dipalmitoyl-sn-glycero-3-phosphocholine (DPPC). Evidence of hydroperoxidation was obtained from the area increase in eosin-containing DOPC and POPC monolayers upon irradiation, which was accompanied by a decrease in monolayer thickness according to grazing incidence X-ray off-specular scattering (GIXOS) data. Furthermore, the changes in polarization-modulated infrared reflection absorption spectroscopy (PM-IRRAS) induced by irradiation were consistent with hydroperoxide migration toward the lipid hydrophilic heads.. In summary, this combination of experimental methods allowed us to determine the effects of eosin Y interaction with cell membrane models under irradiation, which may be associated with the underlying mechanisms of eosin Y as photosensitizer in PDT.

Impact of polyphenol-rich green tea extracts on the protection of DOPC monolayer against damage caused by ozone induced lipid oxidation.

Effectiveness of green tea (compared to two single polyphenols) in removing ozone derived reactive oxygen species acting on dioleoylphosphatidylcholine monolayers was determined. Lipid oxidation was followed by changes in mechanical properties of the layer spread on the aqueous subphase containing various amounts of ozone in the presence and absence of polyphenolics. It was shown that the tea extract (containing 8.5 x 10-4 mg/cm3 polyphenols) is capable of inactivating 0.4 ppm ozone. The DPPH radical scavenging test set polyphenols in the order of increasing activity, consistent with their protective effect in relation to lipid oxidation, showing the highest efficacy of EGCG.

Experimental Modeling of Flavonoid-Biomembrane Interactions.

Nonspecific interactions of flavonoids with lipids can alter the membrane's features (e.g., thickness and fluctuations) as well as influence their therapeutic potentials. However, relatively little is known about the details of how flavonoids interact with lipid components. Structure-dependent interactions of a variety of flavonoids with phospholipid monolayers on a mercury (Hg) film electrode were established by rapid cyclic voltammetry (RCV). The data revealed that flavonoids adopting a planar configuration altered the membrane properties more significantly than nonplanar flavonoids. Quercetin, rutin, and tiliroside were selected for follow-up experiments with Langmuir monolayers, Brewster angle microscopy (BAM), and small-angle X-ray scattering (SAXS). Relaxation phenomena in DOPC monolayers and visualization of the surface with BAM revealed a pronounced monolayer stabilization effect with both quercetin and tiliroside, whereas rutin disrupted the monolayer structure rendering the surface entirely smooth. SAXS showed a monotonous membrane thinning for all compounds studied associated with an increase in the mean fluctuations of the membrane. Rutin, quercetin, and tiliroside decreased the bilayer thickness of DOPC by ∼0.45, 0.8, and 1.1 Å at 6 mol %, respectively. In addition to the novelty of using lipid monolayers to systematically characterize the structure-activity relationship (SAR) of a variety of flavonoids, this is the first report investigating the effect of tiliroside with biomimetic membrane models. All the flavonoids studied are believed to be localized in the lipid/water interface region. Both this localization and the membrane perturbations have implications for their therapeutic activity.

Molecular-Level Modifications Induced by Photo-Oxidation of Lipid Monolayers Interacting with Erythrosin.

Incorporation into cell membranes is key for the action of photosensitizers in photomedicine treatments, with hydroperoxidation as the prominent pathway of lipid oxidation. In this paper, we use Langmuir monolayers of 1,2-dioleoyl-sn-glycero-3-phosphocholine (DOPC) and 1,2-dipalmitoyl-sn-glycero-3-phosphocholine (DPPC) as cell membrane models to investigate adsorption of the photosensitizer erythrosin and its effect on photoinduced lipid oxidation. From surface pressure isotherms and polarization-modulated infrared reflection-absorption spectroscopy (PM-IRRAS) data, erythrosin was found to adsorb mainly via electrostatic interaction with the choline in the head groups of both DOPC and DPPC. It caused larger monolayer expansion in DOPC, with possible penetration into the hydrophobic unsaturated chains, while penetration into the DPPC saturated chains was insignificant. Easier penetration is due to the less packed DOPC monolayer, in comparison to the more compact DPPC according to the monolayer compressibility data. Most importantly, light irradiation at 530 nm made the erythrosin-containing DOPC monolayer become less unstable, with a relative surface area increase of ca. 19%, in agreement with previous findings for bioadhesive giant vesicles. The relative area increase is consistent with hydroperoxidation, supporting the erythrosin penetration into the lipid chains, which favors singlet oxygen generation close to double bonds, an important requirement for photodynamic efficiency.

Interactions of Two Fragments of the Human Antimicrobial Peptide LL-37 with Zwitterionic and Anionic Lipid Monolayers

Abstract The interactions of two fragments of the human antimicrobial LL-37 (LL-32 and LL-20) with lipid monolayers at the soft liquid/air interface have been characterized. To model the interaction with mammalian cell membranes, lipid monolayers composed of the zwitterionic DPPC and DOPC were used. To investigate the interaction with bacterial cell membranes, lipid monolayers of anionic DPPG and POPG were used. DPPC and DPPG exhibit a first-order phase transition from the disordered liquid to the ordered condensed state, whereas POPG and DOPC monolayers are in the fluid disordered state at all surface pressures studied. Therefore, the influence of the monolayer phase state on peptide-lipid interactions can be studied. To obtain insight into the peptide structure and their influence on phospholipid membranes, film balance measurements were coupled with surface sensitive Infrared Reflection-Absorption Spectroscopy (IRRAS). The results were compared to CD measurements in bulk. LL-32 is more surface active and can better intercalate into lipid monolayers than LL-20. Even though LL-32 has no cell-selectivity, our results show how the peptide interacts differently with zwitterionic compared to anionic membrane models. The interaction with DPPC monolayers is based on simple intercalation of the peptides between the lipid molecules. However, the peptides bind in a two-step process to DPPG monolayers, which results in a fluidization of the lipid film. This can be related to a membrane thinning.

Nanoscale Electrostatic Domains in Cholesterol-Laden Lipid Membranes Create a Target for Amyloid Binding

Amyloid fibrils are associated with multiple neurodegenerative disorders, such as Alzheimer’s disease. Although biological membranes are involved in fibril plaque formation, the role of lipid membrane composition in fibril formation and toxicity is not well understood. We investigated the effect of cholesterol on the interaction of model lipid membranes with amyloid-β peptide (Aβ). With atomic force microscopy we demonstrated that binding of Aβ (1–42) to DOPC bilayer, enriched with 20% cholesterol, resulted in an intriguing formation of small nonuniform islands loaded with Aβ. We attribute this effect to the presence of nanoscale electrostatic domains induced by cholesterol in DOPC bilayers. Using frequency-modulated Kelvin probe force microscopy we were able to resolve these nanoscale electrostatic domains in DOPC monolayers. These findings directly affect our understanding of how the presence of cholesterol may induce targeted binding of amyloid deposits to biomembranes. We postulate that this nonhomogeneous electrostatic effect of cholesterol has a fundamental nature and may be present in other lipid membranes and monolayers.