Liquid-ordered Phase Research Articles

Insight into the Exosomal Membrane: From Viewpoints of Membrane Fluidity and Polarity.

Numerous research studies have been done for exosomes, particularly focusing on membrane proteins and included nucleic acids, and the volume of the knowledge about the lipids in the exosomal membrane has been increasing. However, the dynamic property of the exosomal membrane is hardly studied. By employing milk exosome as an example, herein the exosomal membrane was characterized focusing on the membrane fluidity and polarity. The lipid composition and phase state of milk exosome (exosome from bovine milk) were estimated. The milk exosome contained enriched Chol (43.6 mol % in total lipid extracts), which made the membrane in the liquid-ordered (lo) phase by interacting with phospholipids. To suggest a model of exosomal vesicle cargo, the liposome compositions that mimic milk exosome were studied: liposomes were made of cholesterol (Chol), milk sphingomyelin (milk SM), and 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine (POPC). By using fluorescent probes 1,6-diphenyl-1,3,5-hexatriene and 6-dodecanoyl-2-dimethylaminonaphthalene, the microenvironments of submicron-sized membranes of exosome and model liposomes were investigated. The membrane fluidity of milk exosome was slightly higher than those of Chol/milk SM/POPC liposomes with a similar content of Chol, suggesting the presence of enriched unsaturated lipids. The most purposeful membrane property was obtained by the liposome composition of Chol/milk SM/POPC = 40/15/45. From the above, it is concluded that Chol is a fundamental component of the milk exosomal membrane to construct the enriched lo phase, which could increase the membrane rigidity and contribute to the function of exosome.

Sphingomyelinase-Mediated Multitimescale Clustering of Ganglioside GM1 in Heterogeneous Lipid Membranes.

Several signaling processes in the plasma membrane are intensified by ceramides that are formed by sphingomyelinase‐mediated hydrolysis of sphingomyelin. These ceramides trigger clustering of signaling‐related biomolecules, but how they concentrate such biomolecules remains unclear. Here, the spatiotemporal localization of ganglioside GM1, a glycolipid receptor involved in signaling, during sphingomyelinase‐mediated hydrolysis is described. Real‐time visualization of the dynamic remodeling of the heterogeneous lipid membrane that occurs due to sphingomyelinase action is used to examine GM1 clustering, and unexpectedly, it is found that it is more complex than previously thought. Specifically, lipid membranes generate two distinct types of condensed GM1: 1) rapidly formed but short‐lived GM1 clusters that are formed in ceramide‐rich domains nucleated from the liquid‐disordered phase; and 2) late‐onset yet long‐lasting, high‐density GM1 clusters that are formed in the liquid‐ordered phase. These findings suggest that multiple pathways exist in a plasma membrane to synergistically facilitate the rapid amplification and persistence of signals.

Role of cholesterol flip-flop in oxidized lipid bilayers

We performed a series of molecular dynamics simulations of cholesterol (Chol) in nonoxidized 1-palmitoyl-2-linoleoyl-sn-glycero-3-phosphatidylcholine (PLPC) bilayer and in binary mixtures of PLPC-oxidized-lipid-bilayers with 0–50% Chol concentration and oxidized lipids with hydroperoxide and aldehyde oxidized functional groups. From the 60 unbiased molecular dynamics simulations (total of 161 μs), we found that Chol inhibited pore formation in the aldehyde-containing oxidized lipid bilayers at concentrations greater than 11%. For both pure PLPC bilayer and bilayers with hydroperoxide lipids, no pores were observed at any Chol concentration. Furthermore, increasing cholesterol concentration led to a change of phase state from the liquid-disordered to the liquid-ordered phase. This condensing effect of Chol was observed in all systems. Data analysis shows that the addition of Chol results in an increase in bilayer thickness. Interestingly, we observed Chol flip-flop only in the aldehyde-containing lipid bilayer but neither in the PLPC nor the hydroperoxide bilayers. Umbrella-sampling simulations were performed to calculate the translocation free energies and the Chol flip-flop rates. The results show that Chol’s flip-flop rate depends on the lipid bilayer type, and the highest rate are found in aldehyde bilayers. As the main finding, we shown that Chol stabilizes the oxidized lipid bilayer by confining the distribution of the oxidized functional groups.

Ionic environment, thickness and line tension as determinants of phase separation in whole Purified Myelin Membranes monolayers

Purified myelin membranes (PMM) were spread as monomolecular films at the air/aqueous solution interface, and visualized by Brewster Angle Microscopy (BAM) at different lateral pressures (π) on three specific aqueous solutions: absence of salts, physiological conditions and presence of calcium. Coexistence of Liquid-Expanded (LE) and Liquid Ordered (LO) phases persisted up to collapse in the presence of salts, whereas monolayers became homogeneous at π ≥ 35−40 mN/m when salts are absent. This PMM phase-mixing behavior in monolayers is similar to the previously reported behavior of PMM multilamellar vesicles. Reflectivities (Rp) of p-polarized light from both phases were assessed throughout the whole π -range, and film thicknesses (t) were calculated from the Rp values and measured film refractive indices (n). The LO phase was found to be more reflective and thicker than the LE phase at π ≤ 15 mN/m, but less reflective and thinner at higher π.We also determined the line tension (λ) of PMM monolayers at the domain boundaries from the rate of domain shape relaxation, which turned out to be of the order of picoNewtons (pN) and decreased as π increased. A correlation between λ and thickness differences (Δt) was found, suggesting that Δt is a molecular determinant for λ in PMM monolayers.Both λ and Δt were found to increase markedly when calcium was present in the subphase. This result corroborates the concept of divalent cations as a stabilizing factor for phase separation, in line with earlier studies on this mixture forming multilamellar membrane arrangements.

Primary Human Renal Proximal Tubular Epithelial Cells (pHRPTEpiCs): Shiga Toxin (Stx) Glycosphingolipid Receptors, Stx Susceptibility, and Interaction with Membrane Microdomains

Tubular epithelial cells of the human kidney are considered as targets of Shiga toxins (Stxs) in the Stx-mediated pathogenesis of hemolytic–uremic syndrome (HUS) caused by Stx-releasing enterohemorrhagic Escherichia coli (EHEC). Analysis of Stx-binding glycosphingolipids (GSLs) of primary human renal proximal tubular epithelial cells (pHRPTEpiCs) yielded globotriaosylceramide (Gb3Cer) and globotetraosylceramide (Gb4Cer) with Cer (d18:1, C16:0), Cer (d18:1, C22:0), and Cer (d18:1, C24:1/C24:0) as the dominant lipoforms. Investigation of detergent-resistant membranes (DRMs) and nonDRMs, serving as equivalents for the liquid-ordered and liquid-disordered membrane phase, respectively, revealed the prevalence of Gb3Cer and Gb4Cer together with cholesterol and sphingomyelin in DRMs, suggesting lipid raft association. Stx1a and Stx2a exerted strong cellular damage with half-maximal cytotoxic doses (CD50) of 1.31 × 102 pg/mL and 1.66 × 103 pg/mL, respectively, indicating one order of magnitude higher cellular cytotoxicity of Stx1a. Surface acoustic wave (SAW) real-time interaction analysis using biosensor surfaces coated with DRM or nonDRM fractions gave stronger binding capability of Stx1a versus Stx2a that correlated with the lower cytotoxicity of Stx2a. Our study underlines the substantial role of proximal tubular epithelial cells of the human kidney being associated with the development of Stx-mediated HUS at least for Stx1a, while the impact of Stx2a remains somewhat ambiguous.

ESCRT-III induces phase separation in model membranes prior to budding and causes invagination of the liquid-ordered phase

Membrane fission triggered by the endosomal sorting complex required for transport (ESCRT) is an important process observed in several pathogenic and non-pathogenic cellular events. From a synthetic-biology viewpoint, ESCRT proteins represent an interesting machinery for the construction of cell mimetic sub-compartments produced by fission. Since their discovery, the studies on ESCRT-III-mediated action, have mainly focused on protein dynamics, ignoring the role of lipid organization and membrane phase state. Recently, it has been suggested that membrane buds formed by the action of ESCRT-III are generated from transient microdomains in endosomal membranes. However, the interplay between membrane domain formation and ESCRT remodeling pathways has not been investigated. Here, giant unilamellar vesicles made of ternary lipid mixtures, either homogeneous in phase or exhibiting liquid-ordered/liquid-disordered phase coexistence, were employed as a model membrane system. These vesicles were incubated with purified recombinant ESCRT-III proteins from the parasite Entamoeba histolytica. In homogeneous membranes, we observe that EhVps32 can trigger domain formation while EhVps20 preferentially co-localizes in the liquid disordered phase. The addition of EhVps24 appears to induce the formation of intraluminal vesicles produced from the liquid-ordered phase. In phase separated membranes, the intraluminal vesicles are also generated from the liquid-ordered phase and presumably emerge from the phase boundary region. Our findings reinforce the hypothesis that ESCRT-mediated remodeling depends on the membrane phase state. Furthermore, the obtained results point to a potential synthetic biology approach for establishing eukaryotic mimics of artificial cells with microcompartments of specific membrane composition, which can also differ from that of the mother vesicle.

Molecular Mechanism of Ultrasound-Induced Structural Defects in Liposomes: A Nonequilibrium Molecular Dynamics Simulation Study

The use of ultrasound in combination with liposomes is a promising approach to improve drug delivery. To achieve an optimal drug release rate, it is important to understand how ultrasound induces pathways on the liposome surface where drugs can be released from the liposome. To this end, we carry out large-scale ultrasound-induced molecular dynamics simulations for three single lipid component liposomes formed from the commonly used phospholipids: 1,2-dioleoyl-sn-glycero-3-phosphocholine (DOPC), 1,2-dipalmitoylphosphatidylcholine (DPPC), or phosphatidylcholine (POPC). The results show that ultrasound induces the detachment of two leaflets of the DOPC surface, suggesting that the drug release pathway may be through the low lipid packing areas on the stretched surface. In contrast, ultrasound induces pore formation on the surface of DPPC and DOPC, where drugs could escape from the liposomes. While the leaflet detachment and transient pore formation are the mechanisms of DOPC and DPPC, respectively, in both liquid-ordered and liquid-disordered phases, the leaflet detachment mechanism is switched to the transient pore formation mechanism on going from the liquid-ordered phase to the liquid-disordered phase in the POPC liposome. By adding 30% mol cholesterol, the leaflet detachment mechanism is observed in all liposomes. We found that the molecular origin that determines a mechanism is the competition between the intraleaflet and interleaflet interacting energy of lipids. The connection to experimental and theoretical modeling is discussed in some detail.

Enhanced Concanavalin A Binding to Preorganized Mannose Nanoarrays in Glycodendrimersomes Revealed Multivalent Interactions.

The effect of the two‐dimensional glycan display on glycan‐lectin recognition remains poorly understood despite the importance of these interactions in a plethora of cellular processes, in (patho)physiology, as well as its potential for advanced therapeutics. Faced with this challenge we utilized glycodendrimersomes, a type of synthetic vesicles whose membrane mimics the surface of a cell and offers a means to probe the carbohydrate biological activity. These single‐component vesicles were formed by the self‐assembly of sequence‐defined mannose‐Janus dendrimers, which serve as surrogates for glycolipids. Using atomic force microscopy and molecular modeling we demonstrated that even mannose, a monosaccharide, was capable of organizing the sugar moieties into periodic nanoarrays without the need of the formation of liquid‐ordered phases as assumed necessary for rafts. Kinetics studies of Concanavalin A binding revealed that those nanoarrays resulted in a new effective ligand yielding a ten‐fold increase in the kinetic and thermodynamic constant of association.

Morphological and Nanomechanical Properties of Model of Animal and Human Eye Lens Lipid Membranes

The four major phospholipids found in eye lens membrane are phosphatidylcholine (PC), sphingomyelin (SM), phosphatidylethanolamine (PE), and phosphatidylserine (PS). Lens lipid composition of the fiber-cell membrane significantly changes with species. In lower-age animals, phosphatidylcholine (PC) is dominant, whereas, in humans, sphingomyelin (SM) is more dominant. We have used atomic force microscopy (AFM) to study morphological and nanomechanical properties of the lens lipid membrane models from mouse, pig, and humans. Small unilamellar vesicles (SUVs) were prepared from the mixture of four phospholipids (PC, SM, PS, and PE) that resemble mouse, pig, and human phospholipid composition lens membranes. The SUVs phospholipids mixtures were prepared with 0 mol% and 50 mol% of cholesterol. The topographical image and the mechanical properties of animal and human lens model membranes were accessed in a supported lipid membrane (SLM) formed on mica. The topographical images of phospholipid mixtures with 0 mol% cholesterol for both animal and human lens membrane models exhibited coexisting solid-ordered and liquid-disordered phase, where the proportion of solid-ordered phase increases with an increase in SM content in phospholipid mixture. The height difference between the two phases was ∼ 1nm. However, in the presence of 50 mol% of cholesterol, the membrane exhibited only a liquid-ordered phase, both for animal and human phospholipid mixtures. The breakthrough force on the SLM of the animals and human lens model membrane obtained with 50 mol% of cholesterol follow the trends: F (human) > F (pig) > F (mouse). Similarly, the membrane area compressibility modulus (Ka) of the membrane with 50 mol% cholesterol was accessed using the force curve's elastic regime. We found: Ka (human) > Ka (pig) > Ka (mouse). Our results show that phospholipid composition plays a significant role in modulating membrane mechanical properties.

Inter- and Intra-Plane Interactions Control the Existence of Macrodomains in Asymmetric Giant Unilamellar Vesicles

The complexity of the plasma membrane (PM) is in part related to the complexity of its composition. Beyond the hundreds of lipid components in the PM, compositional differences between the exoplasmic and the cytoplasmic leaflets leads to different physical-chemical properties in each leaflet affecting the interaction between them. Model membranes prepared in vitro that reproduce salient properties of the PM can reveal molecular determinants of these interactions. We investigate asymmetric giant unilamellar vesicles (aGUVs) prepared with different lipid compositions by hemifusion. Symmetric GUVs composed of the lipid mixture high-melting lipid (HTm)/DOPC/chol (0.39/0.39/0.22), where HTm saturated lipids ranging from 15-18 carbons, were hemifused with a supported lipid bilayer (SLB) composed of DOPC/chol (0.8/0.2). Fusion of the GUV and SLB outer leaflets promoted a range of lipid exchange, creating asymmetric vesicles. The symmetric GUVs composed of the ternary mixtures exhibited coexistence of liquid-disordered (Ld) and liquid-ordered (Lo) phases. The change in the chain lengths implies different intra-plane interactions in which the domain line tension or the difference in order between the Ld and the Lo phases decreases with the saturated chain length. We found that replacement of the GUV outer leaflet by a composition that forms Ld phase can prevent the formation of macrodomains in a manner that depends on the chain length of the saturated lipid and the percentage of lipid exchange. Interestingly, longer saturated chain length promoted the persistence of macroscopic domains to greater degrees of asymmetry. The domain size transition accompanying changes in the degree of asymmetry can be explained by intra- and inter-leaflet interactions. Our results suggest that cells can control the presence or absence of lipid rafts by controlling the types of lipids present in the two leaflets.

Lipid rafts as platforms for sphingosine 1-phosphate metabolism and signalling

Spontaneous segregation of cholesterol and sphingolipids as a liquid-ordered phase leads to their clustering in selected membrane areas, the lipid rafts. These specialized membrane domains enriched in gangliosides, sphingomyelin, cholesterol and selected proteins involved in signal transduction, organize and determine the function of multiprotein complexes involved in several aspects of signal transduction, thus regulating cell homeostasis. Sphingosine 1-phosphate, an important biologically active mediator, is involved in several signal transduction processes regulating a plethora of cell functions and, not only several of its downstream effectors tend to localize in lipid rafts, some of the enzymes involved in its pathway, of receptors involved in its signalling and its transporters have been often found in these membrane microdomains. Considering this, in this review we address what is currently known regarding the relationship between sphingosine 1-phosphate metabolism and signalling and plasma membrane lipid rafts.

Domain Sorting in Giant Unilamellar Vesicles Adsorbed on Glass.

Giant unilamellar vesicles (GUVs) adsorb to a solid surface and rupture to form a planar bilayer patch. These bilayer patches are used to investigate the properties and functions of biological membranes. Therefore, it is crucial to understand the mechanisms of GUV adsorption. In this study, we investigate the adsorption of phase-separated GUVs on glass using fluorescence microscopy. GUVs containing liquid-ordered (Lo) and liquid-disordered (Ld) phases underwent domain sorting after adsorption. The Ld domain in the unbound region migrated to the highly curved region near the edge of the adsorbed region. Additionally, the Lo phase grew linearly along the edge of the adsorbed region, creating a thin ring-like domain. After the domain sorting event, the GUV ruptured to form a planar bilayer patch with circular-patterned domains in the initially adsorbed area. We found that domain sorting was promoted by increasing the extent of GUV deformation. These results suggest that both the Ld and Lo domains are reorganized for stabilizing the curved bilayer region in adsorbed GUVs.

Role of polyunsaturated phospholipids in liquid-ordered and liquid-disordered phases.

Polyunsaturated phospholipids interact complexly with other membrane components. We have examined pair interactions among ternary lipid bilayers composed of saturated DPPC, polyunsaturated PLePC, and cholesterol in the liquid-ordered and the liquid-disordered phases by all-atom molecular dynamics simulations. The results show that PLePC exhibits strong repulsion with DPPC and cholesterol in the liquid-disordered phase. When the bilayer changes to the liquid-ordered phase, the repulsion of PLePC with DPPC and cholesterol reduces significantly. The phase state of the bilayer which affects the order of acyl tails as well as their density distributions along the bilayer normal is a key factor regulating the role of PLePC in lipid mixtures. Polyunsaturated phospholipids play a strong repulsive role in the liquid-disordered phase but a weak role in the liquid-ordered phase.

Biophysical analysis of the plant-specific GIPC sphingolipids reveals multiple modes of membrane regulation

The plant plasma membrane (PM) is an essential barrier between the cell and the external environment, controlling signal perception and transmission. It consists of an asymmetrical lipid bilayer made up of three different lipid classes: sphingolipids, sterols, and phospholipids. The glycosyl inositol phosphoryl ceramides (GIPCs), representing up to 40% of total sphingolipids, are assumed to be almost exclusively in the outer leaflet of the PM. However, their biological role and properties are poorly defined. In this study, we investigated the role of GIPCs in membrane organization. Because GIPCs are not commercially available, we developed a protocol to extract and isolate GIPC-enriched fractions from eudicots (cauliflower and tobacco) and monocots (leek and rice). Lipidomic analysis confirmed the presence of trihydroxylated long chain bases and 2-hydroxylated very long-chain fatty acids up to 26 carbon atoms. The glycan head groups of the GIPCs from monocots and dicots were analyzed by gas chromatograph–mass spectrometry, revealing different sugar moieties. Multiple biophysics tools, namely Langmuir monolayer, ζ-Potential, light scattering, neutron reflectivity, solid state 2H-NMR, and molecular modeling, were used to investigate the physical properties of the GIPCs, as well as their interaction with free and conjugated phytosterols. We showed that GIPCs increase the thickness and electronegativity of model membranes, interact differentially with the different phytosterols species, and regulate the gel-to-fluid phase transition during temperature variations. These results unveil the multiple roles played by GIPCs in the plant PM.

The C99 domain of the amyloid precursor protein resides in the disordered membrane phase

Processing of the amyloid precursor protein (APP) via the amyloidogenic pathway is associated with the etiology of Alzheimer’s disease. The cleavage of APP by β-secretase to generate the transmembrane 99-residue C-terminal fragment (C99) and subsequent processing of C99 by γ-secretase to yield amyloid-β (Aβ) peptides are essential steps in this pathway. Biochemical evidence suggests that amyloidogenic processing of C99 occurs in cholesterol- and sphingolipid-enriched liquid-ordered phase membrane rafts. However, direct evidence that C99 preferentially associates with these rafts has remained elusive. Here, we tested this by quantifying the affinity of C99-GFP for raft domains in cell-derived giant plasma membrane vesicles (GPMVs). We found that C99 was essentially excluded from ordered domains in vesicles from HeLa cells, undifferentiated SH-SY5Y cells, or SH-SY5Y-derived neurons; instead, ∼90% of C99 partitioned into disordered domains. The strong association of C99 with disordered domains occurred independently of its cholesterol-binding activity or homodimerization, or of the presence of the familial Alzheimer disease Arctic mutation (APP E693G). Finally, through biochemical studies we confirmed previous results, which showed that C99 is processed in the plasma membrane by α-secretase, in addition to the well-known γ-secretase. These findings suggest that C99 itself lacks an intrinsic affinity for raft domains, implying that either i) amyloidogenic processing of the protein occurs in disordered regions of the membrane, ii) processing involves a marginal subpopulation of C99 found in rafts, or iii) as-yet-unidentified protein–protein interactions with C99 in living cells drive this protein into membrane rafts to promote its cleavage therein.

Interaction of drugs with lipid raft membrane domains as a possible target

Introduction: Plasma membranes are not the homogeneous bilayers of uniformly distributed lipids but the lipid complex with laterally separated lipid raft membrane domains, which provide receptor, ion channel and enzyme proteins with a platform. The aim of this article is to review the mechanistic interaction of drugs with membrane lipid rafts and address the question whether drugs induce physicochemical changes in raft-constituting and raft-surrounding membranes.
 Methods: Literature searches of PubMed/MEDLINE and Google Scholar databases from 2000 to 2020 were conducted to include articles published in English in internationally recognized journals. Collected articles were independently reviewed by title, abstract and text for relevance.
 Results: The literature search indicated that pharmacologically diverse drugs interact with raft model membranes and cellular membrane lipid rafts. They could physicochemically modify functional protein-localizing membrane lipid rafts and the membranes surrounding such domains, affecting the raft organizational integrity with the resultant exhibition of pharmacological activity. Raft-acting drugs were characterized as ones to decrease membrane fluidity, induce liquid-ordered phase or order plasma membranes, leading to lipid raft formation; and ones to increase membrane fluidity, induce liquid-disordered phase or reduce phase transition temperature, leading to lipid raft disruption.
 Conclusion: Targeting lipid raft membrane domains would open a new way for drug design and development. Since angiotensin-converting enzyme 2 receptors which are a cell-specific target of and responsible for the cellular entry of novel coronavirus are localized in lipid rafts, agents that specifically disrupt the relevant rafts may be a drug against coronavirus disease 2019.

A Copper Based Catalyst for the CO2 Reduction Electrochemically Synthesized on a Gas Diffusion Layer

The gaseous CO2 concentration in the environment has reached alarming levels1. The uncontrolled exploitation of fossil fuels is one of the main causes for the huge increase of this greenhouse gas and nowadays the living beings have to face the biggest threat to our planet: the Global Warming. In this scenario, a great effort has been demanded to all fields of human sciences in order to guarantee a sustainable future for our society. Among different solutions, the concept of “artificial photosynthesis”2 has gained momentum in view of a smart use of the atmospheric CO2. Inspired by the leaf photosynthesis, the carbon dioxide is converted back to chemicals and fuels by exploiting an electrochemical process3.This contribution describes an easily synthesized catalyst made of metallic copper that can be used for the heterogenous CO2 electroreduction (CO2RR) in liquid phase in order to produce small organic molecules such as formic acid and acetic acid. The metallic copper-based catalyst was electrochemically deposited on a gas diffusion layer (GDL) support made of carbon fibers with a total surface area of 4 cm2. Electrosynthesis allows the one step formation and deposition of the Cu film directly on the carbon paper substrate, whose surface properties were optimised by a pre-treatment in 1M H2SO4 followed by soaking in pure EtOH4. The pre-treatment times were thoroughly evaluated and the ratio between crystalline and amorphous carbon phases was investigated by Raman analysis on the single carbon fibers. Afterwards, as shown in Figure 1, an homogenous electrodeposited Cu film on the carbon fibers support was achieved, starting from an aqueous solution containing Cu(NO3)2 at room temperature and atmospheric pressure. In addition, the reproducibility of the catalyst formation process was tested. The optimized catalyst was employed to carry out the CO2RR in a H-type cell, in which a cation exchange membrane (Nafion-115) divided the cathode side, containing the working electrode, from the anode side where a platinum gauze and Ag/AgCl electrode were used as counter and reference electrodes, respectively. The reaction was performed for one hour in NaHCO3 aqueous solution. At the end of the reaction, all the liquid products were analysed by quantitative H1-NMR, while the total amount of gas products were collected in a gas sampling flask and analysed by Gas Chromatography. As first preliminary tests, several reactions were carried out both in continuous and discontinuous CO2 flux at -1.1 V and -0.7 V vs RHE in which a CO faradaic efficiency (FE) up to 4.36% was obtained with a FE for H2 evolution of 63.80% for the gaseous product. As for the liquid products, formic acid (up to FE = 4.46%) and acetic acid (up to FE = 6.39%) were obtained. References https://www.esrl.noaa.gov/gmd/ccgg/trends/Bensaid S. et al. ChemSusChem 5 (2012) 500 – 521Genovese C. et al. J. Energy Chem. 22 (2013) 202–213 Xuli Ma et al., Int. J. Hydrog. Energy 41(2016) 14553-14561 Figure 1

The interaction of Aβ42 peptide in monomer, oligomer or fibril forms with sphingomyelin/cholesterol/ganglioside bilayers

Aβ42 peptide binds neuronal membranes and aggregates into plaques that are characteristic of Alzheimer's disease. Aβ42 peptide has been proposed to be generated in membrane (nano) domains in the liquid-ordered phase, ganglioside GM1 being a major facilitator of peptide binding to the membrane. The peptide exists in solution in various degrees of aggregation, either monomers, oligomers or fibrils, of which oligomers appear to be particularly toxic. The present study reports on the binding of Aβ42 peptide, in monomer, oligomer or fibril form, to model membranes (lipid vesicles or monolayers), composed of sphingomyelin and cholesterol in equimolar ratios, to which 1–5 mol% of different gangliosides were incorporated. Thermodynamic binding parameters obtained from calorimetric data indicate a strong tendency to bind the membrane (ΔG ≈ 7 kcal/mol peptide), in a process dominated in most cases by the increase in entropy. ΔG was virtually invariant with the ganglioside species and the aggregation state of the peptide. The Langmuir balance demonstrated the capacity of all peptide preparations to become inserted in lipid monolayers of any composition and initial π in the range 10–30 mN/m, although fibrils were less capable to do so than oligomers or monomers, their maximum initial π being ≈25 mN/m.

Preferential Protein Partitioning in Biological Membrane with Coexisting Liquid Ordered and Liquid Disordered Phase Behavior: Underlying Design Principles.

Several studies now show that certain proteins exhibit selective preference toward liquid ordered (L[Formula: see text]) or toward liquid disordered (L[Formula: see text]) regions of the heterogeneous membrane and some of them have preference for the L[Formula: see text]-L[Formula: see text] interface. Spatially heterogenous organization of lipids, enriched in specific protein molecules, function as platforms for signaling and are involved in several other physiologically critical functions. In this review, we collate together some of the experimental observations of cases where proteins preferentially segregate into different phases and highlight the importance of these preferential localization in terms of underlying functions. We also try to understand the structural features and chemical makeup of the membrane-interacting motifs of these proteins. Finally, we put forth some preliminary analysis on class I viral fusion proteins, some of which are known to partition at the L[Formula: see text]-L[Formula: see text] interface, and through them we try to understand the evolutionary design principles of phase segregating proteins. Put together, this review summarizes the existing studies on preferential partitioning of proteins into different membrane phases while emphasizing the need to understand the molecular design-level features that can help us "engineer" functionally rich peptides and proteins with a programmed membrane partitioning.

Single-lipid dynamics in phase-separated supported lipid bilayers

Phase separation is a fundamental organizing mechanism on cellular membranes. Lipid phases have complex dependencies on the membrane composition, curvature, tension, and temperature. Lipid diffusion rates vary by up to ten-fold between liquid-disordered (Ld) and liquid-ordered (Lo) phases depending on the membrane composition, measurement technique, and the surrounding environment. This manuscript reports the lipid diffusion on phase-separated supported lipid bilayers (SLBs) with varying temperature, composition, and lipid phase. Lipid diffusion is measured by single-particle tracking (SPT) and fluorescence correlation spectroscopy (FCS) via custom data acquisition and analysis protocols that apply to diverse membranes systems. Traditionally, SPT is sensitive to diffuser aggregation, whereas the diffusion rates reported by FCS are unaffected by the presence of immobile aggregates. Within this manuscript, we report (1) improved single-particle tracking analysis of lipid diffusion, (2) comparison and consistency between diffusion measurement methods for non-Brownian diffusers, and (3) the application of these methods to measure the phase, temperature, and composition dependencies in lipid diffusion. We demonstrate improved SPT analysis methods that yield consistent FCS and SPT diffusion results even when most fluorescent lipids are frequently confined within aggregates within the membrane. With varying membrane composition and temperature, we demonstrate differences in diffusion between the Ld and Lo phases of SLBs.