Charged Lipids Research Articles

Interaction of Cyanine-D112 with Binary Lipid Mixtures: Molecular Dynamics Simulation and Differential Scanning Calorimetry Study.

This comprehensive molecular dynamics (MD) simulation and experimental study investigates the lipid bilayer interactions of dye D112 for potential photodynamic therapy (PDT) applications. PDT involves formation of a reactive oxidant species in the presence of a light sensitive molecule and light, interrupting cellular functions. D112 was developed as a photographic emulsifier, and we hypothesized that its combined cationic and lipophilic nature can render a superior photosensitizing property–crucial in various light therapies. The focus of this study is to elucidate the binding and insertion mechanisms of D112 with mixed lipid bilayers of anionic dipalmitoyl-phosphatidylserine (DPPS) and zwitterionic dipalmitoyl-phosphatidylcholine (DPPC) lipids to resemble cancer cell membranes. Our studies confirm initial electrostatic binding between the positively charged moieties of D112 and negatively charged lipid headgroups. Additionally, MD simulations combined with differential scanning calorimetry (DSC) studies confirm that D112-lipid interactions are governed by enthalpy-driven nonclassical hydrophobic effects in the membrane interior. It was further noted that despite the electrostatic preference of D112 toward the anionic lipids, D112 molecules colocalized on DPPC-rich domains after insertion. Atomistic level MD studies point toward two possible insertion mechanisms for D112: harpoon and flip. Further insights from the simulation showcase the interactions of low and high aggregates of D112 with the bilayer as the concentration of D112 increases in solution. The size of aggregates modulates the orientation and degree of insertion, providing important information for future studies on membrane permeation mechanisms.

The archaeal division protein CdvB1 assembles into polymers that are depolymerized by CdvC.

The Cdv proteins constitute the cell division system of the Crenarchaea, a machinery closely related to the ESCRT system of eukaryotes. Using a combination of TEM imaging and biochemical assays, we here present an in vitro study of Metallosphaera sedula CdvB1, the Cdv protein that is believed to play a major role in the constricting ring that drives cell division in the Crenarchaea. We show that CdvB1 self‐assembles into filaments that are depolymerized by the Vps4‐homolog ATPase CdvC. Furthermore, we find that CdvB1 binds to negatively charged lipid membranes and can be detached from the membrane by the action of CdvC. Our findings provide novel insight into one of the main components of the archaeal cell division machinery.

A Short Review on Liposome

Liposome, sphere-shaped vesicles consisting of one or more phospholipid bilayer were first described in the 1960’s. Today we are very useful reproduction, reagent and tool in various scientific discipline. Research on liposome technology has progressed from conventional vesicles to ‘second generation’ liposomes in which long-circulating liposomes are obtained by modulating the lipid composition, size and charge on the vesicle.

Coding variants identified in patients with diabetes alter PICK1 BAR domain function in insulin granule biogenesis.

Bin/amphiphysin/Rvs (BAR) domains are positively charged crescent-shaped modules that mediate curvature of negatively charged lipid membranes during remodeling processes. The BAR domain proteins PICK1, ICA69, and the arfaptins have recently been demonstrated to coordinate the budding and formation of immature secretory granules (ISGs) at the trans-Golgi network. Here, we identify 4 coding variants in the PICK1 gene from a whole-exome screening of Danish patients with diabetes that each involve a change in positively charged residues in the PICK1 BAR domain. All 4 coding variants failed to rescue insulin content in INS-1E cells upon knock down of endogenous PICK1. Moreover, 2 variants showed dominant-negative properties. In vitro assays addressing BAR domain function suggested that the coding variants compromised BAR domain function but increased the capacity to cause fission of liposomes. Live confocal microscopy and super-resolution microscopy further revealed that PICK1 resides transiently on ISGs before egress via vesicular budding events. Interestingly, this egress of PICK1 was accelerated in the coding variants. We propose that PICK1 assists in or complements the removal of excess membrane and generic membrane trafficking proteins, and possibly also insulin, from ISGs during the maturation process; and that the coding variants may cause premature budding, possibly explaining their dominant-negative function.

Mimicking Cellular Metabolism in Artificial Cells: Universal Molecule Transport across the Membrane through Vesicle Fusion.

Mass transport across cell membranes is a primary process for cellular metabolism. For this purpose, electrostatically mediated membrane fusion is exploited to transport various small molecules including glucose-6-phosphate, isopropyl β-D-thiogalactoside, and macromolecules such as DNA plasmids from negatively charged large unilamellar vesicles (LUVs) to positively charged giant unilamellar vesicles (GUVs). After membrane fusion between these oppositely charged vesicles, molecules are transported into GUVs to trigger the NAD+ involved enzyme reaction, bacterial gene expression, and in vitro gene expression of green fluorescent protein from a DNA plasmid. The optimized charged lipid percentages are 10% for both positively charged GUVs and negatively charged LUVs to ensure the fusion process. The experimental results demonstrate a universal way for mass transport into the artificial cells through vesicle fusions, which paves a crucial step for the investigation of complicated cellular metabolism.

PFAS fluidize synthetic and bacterial lipid monolayers based on hydrophobicity and lipid charge

Poly- and Perfluoroalkyl substances (PFASs) are pollutants of emerging concern that persist in nature and pose environmental health and safety risks. PFAS disrupt biological membranes resulting in cellular inhibition, but the mechanism of disruption and the role of lipid composition remain unclear. We examine the role of phospholipid saturation and headgroup charge on the interactions between PFASs and phospholipid monolayers comprised of synthetic phosphocholine (PC) and phosphoglycerol (PG) lipids and prepared from bacteria membrane extracts rich in PG lipids from an environmentally relevant marine bacterium Alcanivorax borkumensis. When deposited on a buffered subphase containing PFAS, PFAS mixed within and fluidized zwitterionic and net-anionic monolayers leading to increases in monolayer compressibility that were driven by a combination of PFAS hydrophobicity and monolayer charge density. Differences in the monolayer response using saturated or unsaturated lipids are attributed to the ability of the unsaturated lipids to accommodate PFAS within ‘void space’ arising from the bent lipid tails. Similar fluidization and compressibility behavior were also observed in A. borkumensis lipid monolayers. This work provides new insight into PFAS partitioning into bacterial membranes and the effect PFAS have on the physicomechanical properties of zwitterionic and charged lipid monolayers.

Anionic lipids unlock the gate of pacemaker channels

Hyperpolarization-activated and cyclic nucleotide-modulated channels (HCN) determine pacemaking activity in the heart and brain. Signaling lipids PIP and phosphatidic acid (PA) potentiate HCN activity, but the mechanisms are unknown, since functional studies were performed in cells, with limited control over membrane composition. Here we use SthK, a bacterial homolog of HCN, to investigate lipid-modulation in molecular detail. SthK activity depends on the charge of lipids as revealed by flux-assays and single-channel recordings in defined lipid environments.

Modulation of membrane fusion using a novel fusogenic system composition

Cell membranes are believed to be heterogeneous and to exhibit lipid domains of coexisting phases, which can govern cellular functions and processes. Different factors can affect membrane fusion efficiency, such as the membrane phase state. In this work, we proposed a new composition of the fusogenic system preferentially in the gel phase at room temperature but fluid in physiological temperatures; additionally, membrane fusion efficiency was also investigated. The fusogenic system was based on large unilamellar vesicles (LUVs, 100-150 nm) and composed of an equimolar mixture of neutral and positively charged gel lipids, whereas the GUVs were composed of a mixture of zwitterionic and/or negatively charged fluid lipids.

The effect of lipid charge on phase-separation in ternary lipid membranes

Phase-separation plays an important role in biological membranes as it provides a means to control the spatial organization of molecules on membrane surface and is thus, involved in diverse membrane processes. Prior research on this phenomenon using model membranes has identified a number of ternary lipid compositions that undergo phase separation. These studies have mostly focused on mixtures of cholesterol with neutral lipids such as phosphatidylcholine (PC). Natural membranes are, however, composed of both neutral and charged lipids.

Intramembrane serine protease catalysis in different physical environments

The cleavage of proteins inside cell membranes initiates pathological pathways and is a subject of a major interest. Thus, the nature of the coupling between the physical environment and the function of such proteins has recently attracted significant experimental and theoretical effort. However, it is difficult to determine the nature of this coupling in a unique way by experimental and theoretical studies without separating the chemical and the environmental factors. This work presents calculations of the activation barriers of the intramembrane rhomboid protease in neutral and charged lipid bilayers as well as in detergent micelle, trying to explore the environmental effect.

Metal‐Supported Phospholipid Bilayers Formed by Redox Command (Adv. Mater. Interfaces 6/2022)

A Surface Redox Reaction Lays Down Lipid Membranes In article number 2101289, Ons Hmam and Antonella Badia demonstrate the assembly of single bilayer membranes within minutes at room temperature from phospholipid vesicles in the gel or fluid phase via ion-pairing interactions between the charged lipid head groups and electrogenerated cationic ferroceniums bound to a self-assembled monolayer.

Effects of cholesterol on the size distribution and bending modulus of lipid vesicles.

The influence of cholesterol fraction in the membranes of giant unilamellar vesicles (GUVs) on their size distributions and bending moduli has been investigated. The membranes of GUVs were synthesized by a mixture of two elements: electrically neutral lipid 1, 2-dioleoyl-sn-glycero-3-phosphocholine (DOPC) and cholesterol and also a mixture of three elements: electrically charged lipid 1,2-dioleoyl-sn-glycero-3-phospho-(1′-rac-glycerol) (DOPG), DOPC and cholesterol. The size distributions of GUVs have been presented by a set of histograms. The classical lognormal distribution is well fitted to the histograms, from where the average size of vesicle is obtained. The increase of cholesterol content in the membranes of GUVs increases the average size of vesicles in the population. Using the framework of Helmholtz free energy of the system, the theory developed by us is extended to explain the experimental results. The theory determines the influence of cholesterol on the bending modulus of membranes from the fitting of the proper histograms. The increase of cholesterol in GUVs increases both the average size of vesicles in population and the bending modulus of membranes.

In Vitro Application of Langmuir Monolayer Model: The interfacial Behavior of Myelin Basic Protein with a Plasma Membrane Model.

The stability and compactness of myelin structure and brain homeostasis depend on MBP and glial cell plasma membrane interactions. In order to get more detailed mechanisms of this interaction, the MBP of different concentrations interacting with plasma membrane (POPC/POPE/POPS/Cholesterol (Chol)) model to form bionic membrane was studied by atomic force microscopy (AFM) and Langmuir monolayer technology. The surface pressure(π)-area(A) curve is analyzed by the elastic modulus ([Formula: see text]) and two-dimensional virial equation of state (2D-VES), and the second virial coefficient of the interaction between MBP and plasma membrane molecules was calculated. (i) According to two-dimensional virial equation, it could be analyzed that with the increase of MBP concentration in the subphase, the value of the second virial coefficient increases also, which indicates that MBP is absorbed into lipid membrane of the plasma membrane model at low surface pressure and that the interaction between the molecules is spatial repulsive force, and (ii) in the monolayers with MBP, resulting in an increasing mean molecular area and monolayer stability due to hydrophobic and electrostatic interactions between the positively charged MBP with hydrophobic residues and negatively charged POPS and neutral lipid (POPC, POPE). AFM surface topographic results correspond to the results of the curve analysis, indicating that MBP of different concentrations has significant influences on alignment and conformation of plasma membrane, which is of great medical value and biological significance to the application of interaction between MBP and myelin lipid membrane in treatment of central nervous diseases. Adsorption model of interaction between MBP and plasma membrane model.

Prevalence of polymyxin resistance through the food chain, the global crisis

Antimicrobial resistance is one of the vital challenges facing global health today. Multi-drug resistant (MDR) infections are often treated with the narrow-spectrum drugs, colistin (polymyxin E) or polymyxin B, which are last-resort antibiotics for human therapeutics that are effective against Gram-negative bacteria. Unfortunately, resistance to these polymyxins has occurred because of selective pressure caused by the inappropriate use of those antibiotics, especially in farming. The mechanisms of resistance to polymyxins are mediated through intrinsic, mutational, or genetic alteration in chromosomal genes. The mechanism includes the regulatory network controlling chemical modifications of lipid A moiety of lipopolysaccharide, reducing the negative charge of lipid A and its affinity for polymyxins. Additionally, the unique mobile colistin/polymyxin B resistance (mcr) gene reported in Enterobacteriales is responsible for the horizontal dissemination of resistance to polymyxins via the food chain. There is now an urgent need to increase surveillance for detecting resistance to polymyxins. Therefore, this review presents an overview of presently available scientific literature on the mechanism of resistance to polymyxins, with their associated gene variants, evaluation methods, resistance transmission through the food chain via food bacteria, and related risk factors. We further focus on the significant implications of polymyxins usage in India and future views for food safety to preserve polymyxin activity.

A review on: liposomes

Liposomes, spherical shaped vesicles consisting of one or more phospholipid bilayers, were first described in the mid60s. Today, they are a very useful reproduction, reagent, and tool in various scientific disciplines, including mathematics and theoretical physics, biophysics, chemistry, colloid science, biochemistry, and biology. Among several talented new drug delivery systems, liposomes characterize an advanced technology to deliver active molecules to the site of action. Research on liposome technology has progressed from conventional vesicles to ‘second-generation liposomes’, in which long-circulating liposomes are obtained by modulating the lipid composition, size, and charge of the vesicle. Liposomes with modified surfaces have also been developed using several molecules, such as glycolipids. They are of 0.05- 5.0 micrometer in diameter. Liposomes are used for the treatment of various diseases like tumors or cancer. In this review article provides a Liposomal Drug Delivery System and various aspects related to liposome that can be studied.

Examining the Effect of Charged Lipids on Mitochondrial Outer Membrane Dynamics Using Atomistic Simulations.

The outer mitochondrial membrane (OMM) is involved in multiple cellular functions such as apoptosis, inflammation and signaling via its membrane-associated and -embedded proteins. Despite the central role of the OMM in these vital phenomena, the structure and dynamics of the membrane have regularly been investigated in silico using simple two-component models. Accordingly, the aim was to generate the realistic multi-component model of the OMM and inspect its properties using atomistic molecular dynamics (MD) simulations. All major lipid components, phosphatidylinositol (PI), phosphatidylcholine (PC), phosphatidylethanolamine (PE), and phosphatidylserine (PS), were included in the probed OMM models. Because increased levels of anionic PS lipids have potential effects on schizophrenia and, more specifically, on monoamine oxidase B enzyme activity, the effect of varying the PS concentration was explored. The MD simulations indicate that the complex membrane lipid composition (MLC) behavior is notably different from the two-component PC-PE model. The MLC changes caused relatively minor effects on the membrane structural properties such as membrane thickness or area per lipid; however, notable effects could be seen with the dynamical parameters at the water-membrane interface. Increase of PS levels appears to slow down lateral diffusion of all lipids and, in general, the presence of anionic lipids reduced hydration and slowed down the PE headgroup rotation. In addition, sodium ions could neutralize the membrane surface, when PI was the main anionic component; however, a similar effect was not seen for high PS levels. Based on these results, it is advisable for future studies on the OMM and its protein or ligand partners, especially when wanting to replicate the correct properties on the water-membrane interface, to use models that are sufficiently complex, containing anionic lipid types, PI in particular.

The Electrostatic Basis of Diacylglycerol Pyrophosphate-Protein Interaction.

Diacylglycerol pyrophosphate (DGPP) is an anionic phospholipid formed in plants, yeast, and parasites under multiple stress stimuli. It is synthesized by the phosphorylation action of phosphatidic acid (PA) kinase on phosphatidic acid, a signaling lipid with multifunctional properties. PA functions in the membrane through the interaction of its negatively charged phosphomonoester headgroup with positively charged proteins and ions. DGPP, like PA, can interact electrostatically via the electrostatic-hydrogen bond switch mechanism but differs from PA in its overall charge and shape. The formation of DGPP from PA alters the physicochemical properties as well as the structural dynamics of the membrane. This potentially impacts the molecular and ionic binding of cationic proteins and ions with the DGPP enriched membrane. However, the results of these important interactions in the stress response and in DGPP’s overall intracellular function is unknown. Here, using 31P MAS NMR, we analyze the effect of the interaction of low DGPP concentrations in model membranes with the peptides KALP23 and WALP23, which are flanked by positively charged Lysine and neutral Tryptophan residues, respectively. Our results show a significant effect of KALP23 on the charge of DGPP as compared to WALP23. There was, however, no significant effect on the charge of the phosphomonoester of DGPP due to the interaction with positively charged lipids, dioleoyl trimethylammonium propane (DOTAP) and dioleoyl ethyl-phosphatidylcholine (EtPC). Divalent calcium and magnesium cations induce deprotonation of the DGPP headgroup but showed no noticeable differences on DGPP’s charge. Our results lead to a novel model for DGPP—protein interaction.

Effect of Environmental Factors on the Catalytic Activity of Intramembrane Serine Protease.

The cleavage of protein inside cell membranes regulates pathological pathways and is a subject of major interest. Thus, the nature of the coupling between the physical environment and the function of such proteins has recently attracted significant experimental and theoretical efforts. However, it is difficult to determine the nature of this coupling uniquely by experimental and theoretical studies unless one can separate the chemical and the environmental factors. This work describes calculations of the activation barriers of the intramembrane rhomboid protease in neutral and charged lipid bilayers and in detergent micelle, trying to explore the environmental effect. The calculations of the chemical barrier are done using the empirical valence bond (EVB) method. Additionally, the renormalization method captures the energetics and dynamical effects of the conformational change. The simulations indicate that the physical environment around the rhomboid protease is not a major factor in changing the chemical catalysis and that the conformational and substrate dynamics do not exhibit long-time coupling. General issues about the action of membrane-embedded enzymes are also considered.

Interaction of a short antimicrobial peptide on charged lipid bilayer: A case study on aurein 1.2 peptide

Aurein 1.2 (aurein) is a short but active α-helical antimicrobial peptide discovered in Australian tree frogs (Litoria aurea). It shows inhibition on a broad spectrum of bacteria and cancer cells. With well-defined helicity, amphipathicity, and cationic charges, it readily binds to membranes and causes membrane change and disruption. This study provides details on how aurein interacts with charged lipid membranes by using neutron membrane diffraction (NMD) and neutron spin echo (NSE) spectroscopy on complex peptide-membrane systems. NMD provides higher resolution lipid bilayer structures than solution scattering. NMD revealed the peptide is mostly associated in the lipid headgroup region. Even at moderately high concentrations (e.g., peptide:lipid ratio of 1:30), aurein is located at the acyl chain-headgroup region without deep penetration into the hydrophobic acyl chain. However, it does reduce the elasticity of the membrane at that concentration, which was corroborated by the NSE results. Furthermore, NSE shows that aurein first softens the membrane, like many other α-helical peptides at low concentration, but then makes the membrane much more rigid, even without membrane pore formation. Combining our previous studies, the evidence shows that aurein at relatively low concentrations still modifies lipid distribution significantly and can cause membrane thinning and lateral segregation of charged lipids. At the same time, the membrane's mechanical properties are modified with much slower lipid diffusion. This suggests that aurein can attack the microbial membrane without the need to form membrane pores or disintegrate membranes; instead, it promotes the formation of domains at low concentration.

Analysis of the Interaction between Giant Liposomes and Graphene Oxide

Recently, the interaction between graphene and biological membranes has been under intensive research because graphene and its related materials exhibit distinctive characteristics. For example, water-soluble graphene oxide (GO) is expected to be used as a carrier of drug delivery systems (DDS), but it is also known to have cytotoxicity. Thus, it is important to understand the interaction between lipid bilayers and graphene/GO in order to design molecular systems. Furthermore, these interactions have been studied mostly using small lipid vesicles (< 1 μm), although the size of biological cells is much greater. In this study, we studied the interaction between GO and cell-sized giant unilamellar vesicles (GUVs). We especially examined the effect of the lipid compositions, including neutral, negatively charged, and pyrene-grafted lipids. Pyrene is known to adhere to graphene with the non-covalent π-π stacking interaction. Microscope observations and flow cytometry measurement revealed that GUVs composed of lipids which have attractive interaction with GO rupture at a high probability.