Structure Of Phospholipid Membranes Research Articles

Effects of Antibacterial Peptide F1 on Bacterial Liposome Membrane Integrity.

Previous studies from our lab have shown that the antimicrobial peptide F1 obtained from the milk fermentation by Lactobacillus paracasei FX-6 derived from Tibetan kefir was different from common antimicrobial peptides; specifically, F1 simultaneously inhibited the growth of Gram-negative and Gram-positive bacteria. Here, we present follow-on work demonstrating that after the antimicrobial peptide F1 acts on either Escherichia coli ATCC 25922 (E. coli) or Staphylococcus aureus ATCC 63589 (S. aureus), their respective bacterial membranes were severely deformed. This deformation allowed leakage of potassium and magnesium ions from the bacterial membrane. The interaction between the antimicrobial peptide F1 and the bacterial membrane was further explored by artificially simulating the bacterial phospholipid membranes and then extracting them. The study results indicated that after the antimicrobial peptide F1 interacted with the bacterial membranes caused significant calcein leakage that had been simulated by different liposomes. Furthermore, transmission electron microscopy observations revealed that the phospholipid membrane structure was destroyed and the liposomes presented aggregation and precipitation. Quartz Crystal Microbalance with Dissipation (QCM-D) results showed that the antimicrobial peptide F1 significantly reduced the quality of liposome membrane and increased their viscoelasticity. Based on the study's findings, the phospholipid membrane particle size was significantly increased, indicating that the antimicrobial peptide F1 had a direct effect on the phospholipid membrane. Conclusively, the antimicrobial peptide F1 destroyed the membrane structure of both Gram-negative and Gram-positive bacteria by destroying the shared components of their respective phospholipid membranes which resulted in leakage of cell contents and subsequently cell death.

Antibacterial Mechanism of Multifunctional MXene Nanosheets: Domain Formation and Phase Transition in Lipid Bilayer.

MXenes, two-dimensional metal carbides or nitrides with multifunctional surfaces, are one of the most promising antibacterial nanoscale materials. However, their putative bactericidal mechanism is elusive. To study their bactericidal mechanism, we investigated the interaction between a MXene nanosheet and a model bacterial membrane by molecular dynamics simulations and found that an adsorbed MXene on a membrane surface induced a local phase transition in a domain where the fluidity of the phospholipid in this domain at room temperature was comparable with that of the gel phase. The domain also showed a denser and thinner phospholipid membrane structure than the peripheral phospholipids. By comparing it with our previous experiments of the bactericidal activity of MXenes, we proposed the leakage of intercellular molecules at the phase boundary defects as a possible bactericidal mechanism of MXenes that leads to cell lysis. This study provides a useful model for tailoring new bactericidal nanomaterials.

Trophoblast Stem-Cell-Derived Exosomes Improve Doxorubicin-Induced Dilated Cardiomyopathy by Modulating the let-7i/YAP Pathway

Trophoblast stem cells (TSCs) have been confirmed to play a cardioprotective role in heart failure. However, whether TSC-derived exosomes (TSC-exos) can protect against cardiac injury remains unclear. In the present study, TSC-exos were isolated from the supernatant of TSCs using the ultracentrifugation method and characterized by transmission electron microscopy and western blotting. Utilizing the public Gene Expression Omnibus (GEO) database, we found that let-7i and Yes-associated protein 1 (YAP) could participate in the development of heart failure. In vitro, AC16 cardiomyocytes subjected to doxorubicin (DOX) were treated with TSC-exos or let-7i mimic. Flow cytometry showed that TSC-exos and let-7i both decreased cardiomyocyte apoptosis. In vivo, mice that were intraperitoneally injected into DOX received either PBS, TSC-exos, or AAV9-let7iup for let-7i overexpression. Mice receiving TSC-exos and AAV9-let7iup showed improved cardiac function and decreased inflammatory responses, accompanied by downregulated YAP signaling. Mechanistically, TSC-exos could transfer let-7i to cardiomyocytes and silence the YAP signaling pathway. In conclusion, TSC-exos could alleviate DOX-induced cardiac injury via the let-7i/YAP pathway, which sheds new light on the application of TSC-exos as a potential therapeutic tool for heart failure.

Phospholipid biomarkers in Mars-analogous soils of the Atacama Desert

AbstractIn Mars-analogous hyperarid soils of the Atacama Desert, phospholipid fatty acids (PLFAs) as the main component of biomembrane play a role in reliably determining viable microbes. PLFA analyses illustrated a rise of the microbial abundance (from 5.0 × 106 to 4.2 × 107 cells g−1) and biodiversity (from 7 to 15 different individual PLFAs) from the north hyperarid core of the Atacama Desert to the southern arid region. Abundant cyclopropyl PLFAs (47.2 ± 4.6%) suggested the resistance to oligotrophic and hypersaline environments by Atacama microbial communities. The southernmost arid site had the highest proportion (8.7%) of eukaryotic and fungal lipid biomarkers. Different precipitations (ranging from 0.7 to 2 mm year−1) in the hyperarid core exerted different effects on microbial biomass, PLFA diversity, bacteria and microeukaryotes. By principal component analysis (cumulative 74.6% of variance), the dominance of PLFA hydroxylation was associated with the microbial viability; bacteria rich in C16:0, C18:0 and C16:1ω9 favoured higher soil conductivity and nitrate; and other PLFAs contributed more to the organic content. Additionally by comparing the ratios of PLFAs to well-preserved organics (e.g., mineral-bound organic carbon and microbial phosphorus), I found that the degradation of PLFAs decreased to a minimum when the mean annual precipitation is lower than 2 mm. These findings may further specify identifiable biomarkers on Mars, if potentially extant Martian microbes possess comparable phospholipid membrane structure.

Exosomes as Carriers for Antitumor Therapy.

Exosomes are bilayer vesicles with particle sizes between 50 and 150 nm. Owing to their bilayer membrane structure, cell-to-cell communication, and good absorbability, exosomes are increasingly used as carriers for drug delivery through phospholipid membrane structures to lesion sites with enhanced targeting. Exosome sources and drug-loading methods are important factors affecting their use as drug carriers. There are various ways to pack species in exosomes, and researchers are constantly seeking new and improved approaches. In both in vivo and in vitro evaluations, exosomal vectors have achieved good antitumor efficacies. Despite the importance of exosomes as drug delivery systems with accurate targeting ability and biocompatibility, improvements are needed to facilitate their widespread clinical use. This review focuses on the preparation of exosomes as carriers and their utilization in antitumor research.

The effect of hydronium ions on the structure of phospholipid membranes.

This work seeks to identify the mechanisms by which hydronium ions (H3O+) modulate the structure of phospholipid bilayers by studying the interactions of H3O+ with phospholipids at the molecular level. For this, we carried out multiple microsecond-long unrestrained molecular dynamics (MD) simulations of a POPC bilayer at different H3O+ concentrations. The results show that H3O+ accumulates at the membrane surface where it displaces water and forms strong and long-lived hydrogen bonds with the phosphate and carbonyl oxygens in phospholipids. This results in a concentration-dependent reduction of the area per lipid and an increase in bilayer thickness. This study provides an important molecular-level insight into the mechanism of how H3O+ modulates the structure of biological membranes and is a critical step towards a better understanding of the effect of low pH on mammalian and bacterial membranes.

Membrane destruction and phospholipid extraction by using two-dimensional MoS2 nanosheets.

The interaction of two-dimensional (2D) nanomaterials and bacterial membranes has attracted tremendous attention in antibacterial applications. Various peculiarities of 2D nanomaterials may lead to multiple mechanisms of their interactions with membranes. Here, we investigated the interaction between molybdenum disulfide (MoS2) nanosheets and the bacterial membrane by using both theoretical and experimental approaches. Molecular dynamics simulation presented that MoS2 nanosheets can disrupt the structure of the lipid membrane by making dents on its surface and extracting phospholipid molecules to reduce the integrity of the membrane. This is attributed to the combination of the dispersion interaction of lipid tails with S atoms and the electrostatic interactions of lipid head groups with the Mo and S atoms in the lateral edges of the MoS2 nanosheet. Scanning electron microscopy and transmission electron microscopy confirmed the dents and the destruction of the cell membrane, which would lead to the loss of cytoplasm and the death of bacteria. It should be noted that the phenomenon where MoS2 induces a dent is different from the direct insertion of graphene-based nanomaterials, which might be due to the thicker and stiffer structure of MoS2. Therefore, we believe that the molecular interactions of 2D nanomaterials with bacterial membranes should be highly correlated with their structural characteristics. This newly discovered mechanism of MoS2 nanomaterials to disrupt the cell membrane may promote the application of transition metal dichalcogenide (TMD) nanomaterials in designing remarkable antibacterial materials in the near future.

Structure and phase transitions of DMPC multilamellar vesicles in the presence of Ca2+ ions

Results obtained via small-angle neutron scattering studies of the influence of calcium ions on the structure and phase transitions of phospholipid membranes are presented. The main phase transition temperature of 1,2-dimyristoyl-sn-glycero-3-phosphatidylcholine (1 wt %) multilamellar vesicles is demonstrated to increase by more than 1°C even when the calcium-ion content of the solution is low (0.1 mM). Detailed analysis of the multilamellar vesicles transition between “bound” and “unbound” state indicates the continuous character of the investigated process in both liquid and gel phases. The critical Ca2+ ion concentrations which initiate the destruction of the multilamellar structures and the formation of unilamellar vesicles are found to be ~0.3 mM in the gel and ~0.4–0.5 mM in the liquid-crystal phases during heating and ~0.5 mM in the phases under study upon cooling.

Giant Polymersome Protocells Dock with Virus Particle Mimics via Multivalent Glycan-Lectin Interactions.

Despite the low complexity of their components, several simple physical systems, including microspheres, coacervate droplets and phospholipid membrane structures (liposomes), have been suggested as protocell models. These, however, lack key cellular characteristics, such as the ability to replicate or to dock with extracellular species. Here, we report a simple method for the de novo creation of synthetic cell mimics in the form of giant polymeric vesicles (polymersomes), which are capable of behavior approaching that of living cells. These polymersomes form by self-assembly, under electroformation conditions, of amphiphilic, glycosylated block copolymers in aqueous solution. The glycosylated exterior of the resulting polymeric giant unilamellar vesicles (GUVs) allows their selective interaction with carbohydrate-binding receptor-functionalized particles, in a manner reminiscent of the cell-surface docking of virus particles. We believe that this is the first example of a simple protocell model displaying cell-like behavior through a native receptor-ligand interaction.

Biophysical study of resin acid effects on phospholipid membrane structure and properties

Hydrophobic resin acids (RAs) are synthesized by conifer trees as part of their defense mechanisms. One of the functions of RAs in plant defense is suggested to be the perturbation of the cellular membrane. However, there is a vast diversity of chemical structures within this class of molecules, and there are no clear correlations to the molecular mechanisms behind the RA's toxicity. In this study we unravel the molecular interactions of the three closely related RAs dehydroabietic acid, neoabietic acid, and the synthetic analogue dichlorodehydroabietic acid with dipalmitoylphosphatidylcholine (DPPC) model membranes and the polar lipid extract of soybeans. The complementarity of the biophysical techniques used (NMR, DLS, NR, DSC, Cryo-TEM) allowed correlating changes at the vesicle level with changes at the molecular level and the co-localization of RAs within DPPC monolayer. Effects on DPPC membranes are correlated with the physical chemical properties of the RA and their toxicity.

Oxidative stress in otosclerosis

Objectives: Otosclerosis is a disease involving abnormal bone turnover in the human otic capsule that results in hearing loss. Several hypotheses have been suggested for the etiopathogenesis of otosclerosis; however, its etiology remains unclear.Methods: This study evaluated the correlation between otosclerosis and levels of paraoxonase-1 (PON1), arylesterase, total antioxidant status, total oxidant status (TOS), oxidative stress index (OSI), total sulfhydryl (-SH) groups, lipid hydroperoxide, and ceruloplasmin in the serum of otosclerosis patients and healthy subjects with respect to oxidative stress.Results: In our study, TOS and OSI levels were higher in the otosclerosis patients than in the controls. The PON1 levels showed that oxidative stress was severe, and as a result, antioxidants were consumed and depleted.Discussion: When an imbalance between oxygen free radical production and antioxidative defense mechanisms occurs, reactive oxygen species levels may increase, which in turn may damage cells and tissues through the peroxidation of phospholipid membrane structures. The body initially responds with increased antioxidant production, but if the oxidative stress is severe, decreased antioxidant levels may result. This study reports expression levels of oxidative stress species in otosclerosis patients.

Reconstitution of Adiposome and Artificial Lipid Droplets

The Lipid droplet (LD) is a cellular organelle that consists of a neutral lipid core with a phospholipid monolayer membrane associated with proteins. The presence of this organelle in organisms ranging from bacteria to humans demonstrates that it is highly conserved and may be the first membrane‐bound organelle in living organisms. Recent progress in LD research demonstrates its important roles in energy homeostasis and metabolic syndromes. However, the mechanisms governing its formation, dynamics, interaction with other cellular organelles, and association with proteins have remained elusive. Therefore, we developed an in vitro system to facilitate the elucidation of these mechanisms, based on the reconstitution of artificial LDs. First, we generated sphere‐shaped structures with a neutral lipid core and phospholipid monolayer membrane by mechanically mixing neutral lipids and phospholipids followed by two‐step purification. Since the artificial lipid vesicle, a double layer phospholipid membrane structure, is termed as “liposome”, we named this monolayer phospholipid membrane‐bound structure “adiposome”. We then recruited LD structure‐like/resident proteins to the adiposome, including the bacterial LD protein MLDS, C. elegans LD protein MDT‐28, or mammalian LD protein ADRP/PLIN2. We term the functional protein‐coated adiposomes, “artificial lipid droplets”. Using this experimental system, different proteins can be recruited to build artificial LDs for myriad biological and medical goals.

Label-free characterization of biomembranes: from structure to dynamics

We review recent progress in the study of the structure and dynamics of phospholipid membranes and associated proteins, using novel label-free analytical tools. We describe these techniques and illustrate them with examples highlighting current capabilities and limitations. Recent advances in applying such techniques to biological and model membranes for biophysical studies and biosensing applications are presented, and future prospects are discussed.

Interaction of a trehalose lipid biosurfactant produced by Rhodococcus erythropolis 51T7 with a secretory phospholipase A2

Trehalose-containing glycolipid biosurfactants form an emerging group of interesting compounds, which alter the structure and properties of phospholipid membranes, and interact with enzymatic and non-enzymatic proteins. Phospholipases A2 constitute a class of enzymes that hydrolyze the sn-2 ester of glycerophospholipids, and are classified into secreted phospholipases A2 (sPLA2) and intracellular phospholipases A2. In this work, pancreatic sPLA2 was chosen as a model enzyme to study the effect of the trehalose lipid biosurfactant on enzymes acting on interfaces. By using this enzyme, it is possible to study the modulation of enzyme activity, either by direct interaction of the biosurfactant with the protein, or as a result of the incorporation of the glycolipid on the phospholipid target membrane. It is shown that the succinoyl trehalose lipid isolated from Rhodococcus erythropolis 51T7 interacts with porcine pancreatic sPLA2 and inhibits its catalytic activity. Two modes of inhibition are observed, which are clearly differentiated by its timescale. First, a slow inhibition of sPLA2 activity upon preincubation of the enzyme with trehalose lipid in the absence of substrate is described. Second, incorporation of trehalose lipid into the phospholipid target membrane gives rise to a fast enzyme inhibition. These results are discussed in the light of previous data on sPLA2 inhibitors and extend the list of interesting biological activities reported for this R. erythropolis trehalose lipid biosurfactant.

Phospholipid membrane protection by sugar molecules during dehydration-insights into molecular mechanisms using scattering techniques.

Scattering techniques have played a key role in our understanding of the structure and function of phospholipid membranes. These techniques have been applied widely to study how different molecules (e.g., cholesterol) can affect phospholipid membrane structure. However, there has been much less attention paid to the effects of molecules that remain in the aqueous phase. One important example is the role played by small solutes, particularly sugars, in protecting phospholipid membranes during drying or slow freezing. In this paper, we present new results and a general methodology, which illustrate how contrast variation small angle neutron scattering (SANS) and synchrotron-based X-ray scattering (small angle (SAXS) and wide angle (WAXS)) can be used to quantitatively understand the interactions between solutes and phospholipids. Specifically, we show the assignment of lipid phases with synchrotron SAXS and explain how SANS reveals the exclusion of sugars from the aqueous region in the particular example of hexagonal II phases formed by phospholipids.

Fluorescence microscopic characterization of ionic polymer bead-supported phospholipid bilayer membrane systems

Supported phospholipid membrane structures on cationic organic polymer beads were prepared using mixtures of dioleoylphosphatidylserine (PS) and egg yolk phosphatidylcholine (PC). Confocal fluorescence microscopic observations using a fluorescent membrane probe (N-4-nitrobenzo-2-oxa-1,3-diazole-phosphatidylethanolamine) revealed that the phospholipid molecules in the PS/PC-bead complexes were along the outer surface of the beads, but not inside the beads. The anionic PS on the most outer surface of the PS/PC-bead complexes was responsible for the binding of a positively charged macromolecule, rhodamine isothiocyanate dextran (Mw 70,000) by electrostatic attractive forces. The fluidity of the membranes in the PS/PC-bead complexes was investigated by the fluorescence recovery after a photobleaching technique. The lateral diffusion coefficients (D) for the PS/PC-bead complexes were one-half or less than that for 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine giant unilamellar vesicles without solid supporting materials. Such a constrain of the phospholipid bilayer membrane in the complexes appeared to be due to its immobilization on the cationic polymer bead by electrostatic attractive forces between the PS and ammonium group on the surface of the bead. The D values for the complexes were dependent on the phospholipid composition; the PS(25mol%)/PC(75mol%)-bead complex produced a more fluid membrane than the PS(50mol%)/PC(50mol%)-bead one. Thus, the fluidity of the phospholipid bilayer membranes formed on the cationic polymer beads was significantly affected by the anionic phospholipid fraction used for the preparation of the complexes.

Metabolic gray matter changes of adolescents with anorexia nervosa in combined MR proton and phosphorus spectroscopy

There are hints for changes in phospholipid membrane metabolism and structure in the brain of adolescents with anorexia nervosa (AN) using either proton ((1)H) or phosphorus ((31)P) magnetic resonance spectroscopic imaging (MRSI). We aimed to specify these pathological metabolite changes by combining both methods with additional focus on the neuronal metabolites glutamate (Glu) and N-acetyl-l-aspartate (NAA). Twenty-one female patients (mean 14.4 ± 1.9 years) and 29 female controls (mean 16 ± 1.6 years) underwent (1)H and (31)P MRSI at 3 T applied to the centrum semiovale including the anterior cingulate cortex. We assessed gray matter (GM) and white matter (WM) metabolite concentration changes of the frontal and parietal brain measuring choline(Cho)- and ethanolamine(Eth)-containing compounds, Glutamate (Glu) and glutamine (Gln) and their sum (Glx), myoinositol, NAA, and high-energy phosphates. For (1)H MRSI, a clear discrimination between GM and WM concentrations was possible, showing an increase of Glx (p < 0.001), NAA (frontal p < 0.05), pooled creatine (tCr) (p < 0.001), and choline (tCho) (p < 0.05) in the GM of AN patients. The lipid catabolites glycerophosphocholine (p < 0.07) and glycerophosphoethanolamine (p < 0.03) were increased in the parietal region. Significant changes in GM metabolite concentrations were observed in AN possibly triggered by elevated excitotoxin Glu. Increased tCho may indicate modifications of membrane phospholipids due to increased catabolism in the parietal region. Since no significant changes in phosphorylated choline compounds were found for the frontal region, the tCho increase in this region may hint to fluidity changes.

Force Spectroscopy Reveals the Effect of Different Ions in the Nanomechanical Behavior of Phospholipid Model Membranes: The Case of Potassium Cation

How do metal cations affect the stability and structure of phospholipid bilayers? What role does ion binding play in the insertion of proteins and the overall mechanical stability of biological membranes? Investigators have used different theoretical and microscopic approaches to study the mechanical properties of lipid bilayers. Although they are crucial for such studies, molecular-dynamics simulations cannot yet span the complexity of biological membranes. In addition, there are still some experimental difficulties when it comes to testing the ion binding to lipid bilayers in an accurate way. Hence, there is a need to establish a new approach from the perspective of the nanometric scale, where most of the specific molecular phenomena take place. Atomic force microscopy has become an essential tool for examining the structure and behavior of lipid bilayers. In this work, we used force spectroscopy to quantitatively characterize nanomechanical resistance as a function of the electrolyte composition by means of a reliable molecular fingerprint that reveals itself as a repetitive jump in the approaching force curve. By systematically probing a set of bilayers of different composition immersed in electrolytes composed of a variety of monovalent and divalent metal cations, we were able to obtain a wealth of information showing that each ion makes an independent and important contribution to the gross mechanical resistance and its plastic properties. This work addresses the need to assess the effects of different ions on the structure of phospholipid membranes, and opens new avenues for characterizing the (nano)mechanical stability of membranes.

Microwave induced shift of the main phase transition in phosphatidylcholine membranes

Numerous experimental evidence show that exposure of biological systems to extremely high frequency microwaves may induce significant effects even at low powers. These effects are thought to occur via nonthermal mechanisms involving primarily the interaction of microwaves with phospholipid membrane structures. However, no conclusive experimental evidence that biomembranes exhibit remarkable sensitivity to this radiation has been provided up to now. Here, deuterium nuclear magnetic resonance spectroscopy is used to study the effects of microwaves on 1,2-Dimyristoyl-sn-glycero-3-phosphatidylcholine/ 2H 2O multilamellar vesicles that serve as biomimetic membranes. Here we show that, if the membrane is brought into close proximity to the transition point, microwaves induce a reduction of water ordering at the membrane interface, an upward shift of the main phase transition temperature and a broadening of the transition region. A deep dosimetric analysis shows that the above effects are nonthermal, indicating the need for a nonthermal hypothesis to explain them. This study suggests that exposure to high-frequency microwaves can have far reaching consequences on active biological systems.

Alzheimer's disease amyloid- β peptide analogue alters the ps-dynamics of phospholipid membranes

We have investigated the influence of the neurotoxic Alzheimer's disease peptide amyloid- β (25–35) on the dynamics of phospholipid membranes by means of quasi-elastic neutron scattering in the picosecond time-scale. Samples of pure phospholipids (DMPC/DMPS) and samples with amyloid- β (25–35) peptide included have been compared. With two different orientations of the samples the directional dependence of the dynamics was probed. The sample temperature was varied between 290 K and 320 K to cover both the gel phase and the liquid-crystalline phase of the lipid membranes. The model for describing the dynamics combines a long-range translational diffusion of the lipid molecules and a spatially restricted diffusive motion. Amyloid- β (25–35) peptide affects significantly the ps-dynamics of oriented lipid membranes in different ways. It accelerates the lateral diffusion especially in the liquid-crystalline phase. This is very important for all kinds of protein–protein interactions which are enabled and strongly influenced by the lateral diffusion such as signal and energy transducing cascades. Amyloid- β (25–35) peptide also increases the local lipid mobility as probed by variations of the vibrational motions with a larger effect in the out-of-plane direction. Thus, the insertion of amyloid- β (25–35) peptide changes not only the structure of phospholipid membranes as previously demonstrated by us employing neutron diffraction (disordering effect on the mosaicity of the lipid bilayer system) but also the dynamics inside the membranes. The amyloid- β (25–35) peptide induced membrane alteration even at only 3 mol% might be involved in the pathology of Alzheimer's disease as well as be a clue in early diagnosis and therapy.