The large number of potential applications of ionic liquids (ILs) requires an understanding of their environmental impacts including their adverse effects on microorganisms living in soil and water. The molecular mechanism of toxic activities of these liquids is yet to be understood in detail. Any foreign molecules, interacting with an organism, have to encounter first the cellular membrane, which is predominantly composed of the lipid bilayer. In this work, multilamellar vesicles (MLV) of phospholipids have been used to shed light on the effect of an IL on the structure of a cellular membrane. The MLVs formed by the zwitterionic lipid, 1,2-dipalmitoyl-sn-glycero-3-phosphocholine (DPPC) are found to shrink as a consequence of interaction with an imidazolium-based IL, 1-decyl-3-methylimidazolium tetrafluoroborate ([DMIM] [BF 4 ]). The absorbed ILs significantly modify the surface charge of the MLVs. While these observations indicate a strong membrane-IL interaction, synchrotron-based small angle X-ray diffraction (SAXD) measurements provide a structural description of the interaction. SAXD and Fourier transform infrared spectroscopy studies clearly reveal a disordering effect of the IL on the conformational organization of the lipid chains. The presence of the negatively charged lipid 1,2-dipalmitoyl-sn-glycero-3-phospho-l-serine sodium salt (DPPS) in MLVs plays an important role in disordering the chains in the membrane and inter-bilayer interactions. © 2018, European Biophysical Societies' Association.

Sajal Kumar Ghosh

Physics

(SoNS) School of Natural Sciences

Mitra S.

Ray D.

Bhattacharya G.

Gupta R.

Sen D.

Aswal V.K.

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Shiv Nadar University

European Biophysics Journal

Ionic liquids (ILs) are the attractions of researchers today due to their vast area of potential applications. For biomedical uses, it becomes essential to understand their interactions with cellular membrane. Here, the membrane is mimicked with lipid bilayer and monolayer composed of liver lipids extract. Three archetypal imidazolium based ILs, 1-decyl-3-methylimidazolium tetrafluoroborate ([DMIM][BF4] or [C10MIM][BF4]), 1-octyl-3-methylimidazolium tetrafluoroborate, ([OMIM][BF4] or [C8MIM][BF4]) and 1-ethyl-3-methylimidazolium tetrafluoroborate ([EMIM][BF4] or [C2MIM][BF4]) having different alkyl chain lengths are used in the present study. The isothermal titration calorimetry (ITC) measurements showed that [DMIM][BF4] interacts strongest with the liver lipid membrane compared to other two ILs which have relatively shorter alkyl chain length. The low values of stoichiometry ratio of ILs indicates that ILs penetrate within the core of the lipid bilayer. The interaction of ILs with the liver lipid membrane is found to be mainly driven by entropy which could be due to the change in the structure of the lipid membrane at local or global scales. Dynamic light scattering (DLS) measurements indicate that there are no changes in the size of vesicles due to addition of [DMIM][BF4] indicating stability of the vesicles. On the other hand, x-ray reflectivity (XRR) measurements showed a concentration dependent change in the monolayer structure. At low concentration of the IL, the monolayer thickness decreases, exhibiting an increase in the electron density of the layer. However, at higher concentrations, the monolayer thickness increases proving a concentration dependent effects of the IL on the arrangement of the molecules. © 2021

Biochimica et Biophysica Acta - Biomembranes

Thermodynamics and structure of model bio-membrane of liver lipids in presence of imidazolium-based ionic liquids

A cellular membrane, primarily a lipid bilayer, surrounds the internal components of a biological cell from the external components. This self-assembled bilayer is known to be perturbed by ionic liquids (ILs) causing malfunctioning of a cellular organism. In the present study, surface-sensitive X-ray scattering techniques have been employed to understand this structural perturbation in a lipid multilayer system formed by a zwitterionic phospholipid, 1,2-dipalmitoyl-sn-glycero-3-phosphocholine. The ammonium and phosphonium-based ILs with methanesulfonate anions are observed to induce phase-separated domains in the plane of a bilayer. The lamellar X-ray diffraction peaks suggest these domains to correlate across the bilayers in a smectic liquid crystalline phase. This induced IL-rich lamellar phase has a very low lamellar repeat distance, suggesting the formation of an interdigitated bilayer. The IL-poor phase closely related to the pristine lipid phase shows a decrement in the in-plane chain lattice parameters with a reduced tilt angle. The ammonium and phosphonium-based ILs with a relatively bulky anion, p-toluenemethanesulfonate, have shown a similar effect. © 2021 American Chemical Society. All rights reserved.

Fulltext

ACS Omega

Ionic liquid-induced phase-separated domains in lipid multilayers probed by x-ray scattering studies

Abstract.: The physical properties of an aqueous solution of a macromolecule primarily depend on its chemical structure and the mesoscopic aggregates formed by many of such molecules. Ionic liquids (ILs) are the macromolecules that have caught significant research interests for their enormous industrial and biomedical applications. In the present paper, the physical properties, such as density, viscosity, ionic conductivity of aqueous solutions of various ILs, have been investigated. These properties are found to systematically depend on the shape and size of the anion and the cation along with the solution concentration. The ionic conductivity and viscosity behavior of the solutions do not strictly follow the Walden rule that relates the conductivity to the viscosity of the solution. However, the modified Walden rule could explain the behavior. A simple calculation based on the geometry of a given molecule could shed the light on the observed results. Graphical abstract: [Figure not available: see fulltext.] © 2020, EDP Sciences, Società Italiana di Fisica and Springer-Verlag GmbH Germany, part of Springer Nature.

European Physical Journal E

Relating the physical properties of aqueous solutions of ionic liquids with their chemical structures

The potential biomedical applications of ionic liquids (ILs) need an understanding of the effects of the molecules in living organisms. Researchers are still exploring the molecular level description of this interaction. Any foreign molecule affecting a living organism has to interact first with the outer layer of a cell, which is mainly composed of a phospholipid bilayer. Here, to understand the membrane-IL interaction, a zwitterionic lipid has been chosen to prepare multilamellar vesicles (MLVs) which are the model cellular membranes. Small angle x-ray diffraction study has revealed the change in inter-bilayer spacing in the presence of imidazolium based ILs in the ripple phase of the bilayer. The investigation was extended further to understand the role of a negatively charged lipid in the MLVs. This study indicates that the cations of an IL screen electrostatic repulsion between the negatively charged lipids and induce the formation of a vesicle. © 2020 American Institute of Physics Inc.. All rights reserved.

AIP Conference Proceedings

Effect of ionic liquids on the structures of ripple phases of model cellular membranes

There are great efforts of synthesizing imidazolium-based ionic liquids (ILs) for developing new antibiotics as these molecules have shown strong antibacterial activities. Compared to a single-hydrocarbon-chained IL, the lipid analogues (LAs) with two chains are more effective. In the present study, the LA molecule MeIm(COOH)Me(Oleylamine)Iodide has been synthesized and its surface activities along with the effectiveness in restructuring of a model cellular membrane have been quantified. The molecule is found to be highly surface active as estimated from the area-pressure isotherm of a monolayer of the molecules formed at the air-water interface. The X-ray reflectivity (XRR) studies of a monolayer dip-coated on a hydrophilic substrate have shown the structural properties of the layer which resembles to those of unsaturated phospholipids. The LA molecules are observed to fluidize a phospholipid bilayer formed by the saturated lipid 1,2-dipalmitoyl-sn-glycero-3-phosphocholine (DPPC). At a lower surface pressure, the lipid monolayer of DPPC has exhibited a thickening effect at a low concentration of added LA and a thinning effect at higher concentration. However, at a high surface pressure of the monolayer, the thickness is found to decrease monotonically. The in-plane pressure-dependent interaction of LA molecules with model cellular membrane and the corresponding perturbation in the structure and physical properties of the membrane may be linked to the strong lysing effect of these types of molecules. © 2019 American Chemical Society.

Langmuir

Surface Activities of a Lipid Analogue Room-Temperature Ionic Liquid and Its Effects on Phospholipid Membrane

Ionic liquids (ILs) have generated considerable attention recently because of their cytotoxicity and application as antibiotics. However, the mechanism of how they damage cell membranes is not currently well understood. In this paper, the antibacterial activities of two imidazolium-based ILs, namely 1-butyl- 3-methylimidazolium tetrafluroborate ([BMIM][BF4]) and 1-ethyl- 3-methylimidazolium tetrafluroborate ([EMIM][BF4]) have been investigated. The activity of [BMIM][BF4] on gram negative bacteria E. coli is observed to be stronger compared with the short chained [EMIM][BF4]. To explain this observation, the effects of these ILs on the self-assembled structures of model cellular membranes have been investigated. The in-plane elasticity of a monolayer formed at air-water interface by 1,2-dipalmitoyl- sn-glycero- 3-phosphocholine (DPPC) lipids was reduced in the presence of the ILs. The x-ray reflectivity studies on polymer supported lipid bilayer have shown the bilayer to shrink and correspondingly exhibit an increase in electron density. The presence of a certain mol\% of negatively charged lipid, 1,2-dipalmitoyl-rac-glycero-3-phospho-L-serine (DPPS), in DPPC mono- and bi-layers enhances the effect considerably. © 2018 Elsevier B.V.

Chemistry and Physics of Lipids

Structural changes in cellular membranes induced by ionic liquids: From model to bacterial membranes

Understanding the interaction of ionic liquids with cellular membrane becomes utterly important to comprehend the activities of these liquids in living organisms. Lipid monolayer formed at the air–water interface is employed as a model system to follow this interaction by investigating important thermodynamic parameters. The penetration kinetics of the imidazolium-based ionic liquid 1-decyl-3-methylimidazolium tetrafluoroborate ([DMIM][BF4]) into the zwitterionic 1,2-dipalmitoyl-sn-glycero-3-phosphocholine (DPPC) lipid layer is found to follow the Boltzmann-like equation that reveals the characteristic time constant which is observed to be the function of initial surface pressure. The enthalpy and entropy calculated from temperature-dependent pressure–area isotherms of the monolayer show that the added ionic liquids bring about a disordering effect in the lipid film. The change in Gibbs free energy indicates that an ionic liquid with longer chain has a far greater disordering effect compared to an ionic liquid with shorter chain. The differential scanning calorimetric measurement on a multilamellar vesicle system shows the main phase transition temperature to shift to a lower value, which, again, indicates the disordering effect of the ionic liquid on lipid membrane. All these studies fundamentally point out that, when ionic liquids interact with lipid molecules, the self-assembled structure of a cellular membrane gets perturbed, which may be the mechanism of these molecules having adverse effects on living organisms. © 2018, International Union for Pure and Applied Biophysics (IUPAB) and Springer-Verlag GmbH Germany, part of Springer Nature.

Biophysical Reviews

Thermodynamics of interaction of ionic liquids with lipid monolayer

The interaction of phospholipids with the peripheral membrane proteins like spectrin is important not only to understand the various physiological functions of cells, but also to gain insight into the mechanism involved in the self-assembly of polymer-like long chain molecules at the soft surfaces and interfaces. The lipid head-group specificity of adsorption of spectrin to supported phopsholipid bilayer model membranes has been investigated using the X-ray reflectivity (XRR) technique. Model lipid bilayers composed of phosphatidylcholine (PC) and phosphatidylethanolamine (PE) head groups have been prepared on a soft polymer cushion and the XRR measurements have been carried out from the bilayers immersed in a water bath using high-energy synchrotron X-rays. Our results suggest that in PC-based membranes the spectrin chains form a uniform layer on top of the bilayer with their chains lying on the membrane surface, while in PE-based membranes with relatively smaller head groups, the spectrin chains are attached only through a few possible binding sites with the rest of the part projected out of the membrane surface. In addition, the reflectivity profiles reveal the penetration of spectrin polypeptide chains through the PE bilayer in its fluid phase. Pressure-area isotherm measurements on Langmuir monolayers also support similar observations on the adsorption of spectrin molecules to the membranes composed of PC and PE. The observed results were explained using a qualitative model based on the ion-mediated protein interaction in the PC-based membrane. © CopyrightEPLA, 2017.

Differential adsorption of a membrane skeletal protein, spectrin, in phospholipid membranes

Ionic liquids (ILs) are important for their antimicrobial activity and are found to be toxic to some microorganisms. To shed light on the mechanism of their activities, the interaction of an imidazolium-based IL 1-butyl-3-methylimidazolium tetrfluoroborate ([BMIM][BF4]) with E. coli bacteria and cell-membrane-mimicking lipid mono- and bilayers has been studied. The survival of the bacteria and corresponding growth inhibition are observed to be functions of the concentration of the IL. The IL alters the pressure-area isotherm of the monolayer formed at an air-water interface by the 1,2-dipalmitoyl-sn-glycero-3-phosphocholine (DPPC) lipid. The in-plane elasticity of the lipid layer is reduced as a consequence of the insertion of this IL. The X-ray reflectivity study from a polymer-supported lipid bilayer shows strong perturbation in the self-assembled structure of the bilayer due to the interaction. As a consequence, there is a considerable decrease in bilayer thickness and a corresponding increase in electron density. These results, however, depend on the chain configurations of the lipid molecules. © 2017 American Chemical Society.

X-ray Reflectivity Study of the Interaction of an Imidazolium-Based Ionic Liquid with a Soft Supported Lipid Membrane

Ionic liquids (ILs) are potential candidates for new antimicrobials due to their tunable antibacterial and antifungal properties that are required to keep pace with the growing challenge of bacterial resistance. To a great extent their antimicrobial actions are related to the interactions of ILs with cell membranes. Here, we report the effects of ILs on the nanoscopic dynamics and phase behaviour of a dimyristoylphosphatidylcholine (DMPC) membrane, a model cell membrane, as studied using neutron scattering techniques. Two prototypical imidazolium-based ILs 1-butyl-3-methylimidazolium tetrafluoroborate (BMIM[BF4]) and 1-decyl-3-methylimidazolium tetrafluoroborate (DMIM[BF4]), which differ only in terms of the alkyl chain length of cations, have been used for the present study. Fixed Elastic Window Scan (FEWS) shows that the incorporation of ILs affects the phase behaviour of the phospholipid membrane significantly and the transition from a solid gel to a fluid phase shifts to lower temperature. This is found to be consistent with our differential scanning calorimetry measurements. DMIM[BF4], which has a longer alkyl chain cation, affects the phase behaviour more strongly in comparison to BMIM[BF4]. The pressure-area isotherms of the DMPC monolayer measured at the air-water interface show that in the presence of ILs, isotherms shift towards higher area-per lipid molecule. DMIM[BF4] is found to shift the isotherm to a greater extent compared to BMIM[BF4]. Quasielastic neutron scattering (QENS) data show that both ILs act as a plasticizer, which enhances the fluidity of the membrane. DMIM[BF4] is found to be a stronger plasticizing agent in comparison to BMIM[BF4] that has a cation with a shorter alkyl chain. The incorporation of DMIM[BF4] enhances not only the long range lateral motion but also the localised internal motion of the lipids. On the other hand, BMIM[BF4] acts weakly in comparison to DMIM[BF4] and mainly alters the localised internal motion of the lipids. Any subtle change in the dynamical properties of the membrane can profoundly affect the stability of the cell. Hence, the dominant effect of the IL with the longer chain length on the dynamics of the phospholipid membrane might be correlated with its cytotoxic activity. QENS data analysis has provided a quantitative description of the effects of the two imidazolium-based ILs on the dynamical and phase behaviour of the model cell membrane, which is essential for a detailed understanding of their action mechanism. © 2017 The Royal Society of Chemistry.