Supported Lipid Membranes Research Articles

Variable Non-Gaussian Transport of Nanoplastic on Supported Lipid Bilayers in Saline Conditions.

Nanoplastic-lipid interaction is vital to understanding the nanoscale mechanism of plastic adsorption and aggregation on a lipid membrane surface. However, a single-particle mechanistic picture of the nanoplastic transport process on a lipid surface remains unclear. Here, we report a salt-dependent non-Gaussian transport mechanism of polystyrene particles on a supported 1-palmitoyl-2-oleoyl-glycero-3-phosphocholine (POPC) lipid bilayer surface. Particle stickiness on the POPC surface increases with salt concentration, where the particles stay longer at the surface and diffuse to shorter distances. Additionally, a non-Gaussian diffusion state dominates the transport process at high salt concentrations. Our current study provides insight into the transport mechanism of polystyrene (PS) particles on supported lipid membranes, which is essential to understanding fundamental questions regarding the adsorption mechanisms of nanoplastics on lipid surfaces.

Unraveling the Biophysical Mechanisms of How Antiviral Detergents Disrupt Supported Lipid Membranes: Toward Replacing Triton X-100.

Triton X-100 (TX-100) is a membrane-disrupting detergent that is widely used to inactivate membrane-enveloped viral pathogens, yet is being phased out due to environmental safety concerns. Intense efforts are underway to discover regulatory acceptable detergents to replace TX-100, but there is scarce mechanistic understanding about how these other detergents disrupt phospholipid membranes and hence which ones are suitable to replace TX-100 from a biophysical interaction perspective. Herein, using the quartz crystal microbalance-dissipation (QCM-D) and electrochemical impedance spectroscopy (EIS) techniques in combination with supported lipid membrane platforms, we characterized the membrane-disruptive properties of a panel of TX-100 replacement candidates with varying antiviral activities and identified two distinct classes of membrane-interacting detergents with different critical micelle concentration (CMC) dependencies and biophysical mechanisms. While all tested detergents formed micelles, only a subset of the detergents caused CMC-dependent membrane solubilization similarly to that of TX-100, whereas other detergents adsorbed irreversibly to lipid membrane interfaces in a CMC-independent manner. We compared these biophysical results to virus inactivation data, which led us to identify that certain membrane-interaction profiles contribute to greater antiviral activity and such insights can help with the discovery and validation of antiviral detergents to replace TX-100.

Nanoscale visualization of phase separation in binary supported lipid monolayer using tip-enhanced Raman spectroscopy.

Supported lipid membranes are an important model system to study the phase separation behavior at the nanoscale. However, the conventional nanoanalytical tools often fail to provide reliable chemical characterization of the phase separated domains in a non-destructive and label-free manner. This study demonstrates the application of scanning tunneling microscopy-based tip-enhanced Raman spectroscopy (TERS) to study the nanoscale phase separation in supported d62-DPPC : DOPC lipid monolayers. Hyperspectral TERS imaging successfully revealed a clear segregation of the d62-DPPC-rich and DOPC-rich domains. Interestingly, nanoscale deposits of d62-DPPC were observed inside the DOPC-rich domains and vice versa. High-resolution TERS imaging also revealed the presence of a 40-120 nm wide interfacial region between the d62-DPPC-rich and DOPC-rich domains signifying a smooth transition rather than a sharp boundary between them. The novel insights obtained in this study demonstrate the effectiveness of TERS in studying binary lipid monolayers at the nanoscale.

Structure and thermodynamics of supported lipid membranes on hydrophobic van der Waals surfaces.

Understanding the adsorption and physical characteristics of supported lipid membranes is crucial for their effective use as model cell membranes. Their morphological and thermodynamic properties at the nanoscale have traditionally been studied on hydrophilic substrates, such as mica and silicon oxide, which have proved to facilitate the reconstruction of biomembranes. However, in more recent years, with the advent of the van der Waals crystals technology, two-dimensional crystals such as graphene have been proposed as potential substrates in biosensing devices. Membranes formed on these crystals are expected to behave differently owing to their intrinsic hydrophobicity, however thus far knowledge of their morphological and thermodynamic properties is lacking. Here we present a comprehensive nanoscale analysis of the adsorption of phosphatidylcholine lipid monolayers on two of the most commonly used van der Waals crystals, graphite and hexagonal boron nitride. Both morphological and thermodynamic properties of the lipid membranes were investigated using temperature-controlled atomic force microscopy. Our experiments show that the lipids adsorb onto the crystals, forming monolayers with their orientation dependent upon their concentration. Furthermore, we found that the hydrophobicity of van der Waals crystals determines a strong increase in the transition temperature of the lipid monolayer compared to that observed on hydrophilic substrates. These results are important for understanding the properties of lipid membranes at solid surfaces and extending their use to novel drug delivery and biosensing devices made of van der Waals crystals.

Atomic-scale structure of interfacial water on gel and liquid phase lipid membranes.

Hydration of biological membranes is essential to a wide range of biological processes. In particular, it is intrinsically linked to lipid thermodynamic properties, which in turn influence key cell functions such as ion permeation and protein mobility. Experimental and theoretical studies of the surface of biomembranes have revealed the presence of an interfacial repulsive force, which has been linked to hydration or steric effects. Here, we directly characterise the atomic-scale structure of water near supported lipid membranes of 1,2-dimyristoyl-sn-glycero-3-phosphocholine in their gel and liquid phase through three-dimensional atomic force microscopy (3D AFM). First, we demonstrate the ability to probe the morphology of interfacial water of lipid bilayers in both phases with sub-molecular resolution by using ultrasharp tips. We then visualise the molecular arrangement of water at the lipid surface at different temperatures. Our experiments reveal that water is organised in multiple hydration layers on both the solid-ordered and liquid-disordered lipid phases. Furthermore, we observe a monotonic repulsive force, which becomes relevant only in the liquid phase. These results offer new insights into the water structuring near soft biological surfaces, and demonstrate the importance of investigating it with vertical and lateral sub-molecular resolution.

Lipid membranes supported by polydimethylsiloxane substrates with designed geometry.

The membrane curvature of cells and intracellular compartments continuously adapts to enable cells to perform vital functions, from cell division to signal trafficking. Understanding how membrane geometry affects these processes in vivo is challenging because of the biochemical and geometrical complexity as well as the short time and small length scales involved in cellular processes. By contrast, in vitro model membranes with engineered curvature would provide a versatile platform for this investigation and applications to biosensing and biocomputing. Here, we present a strategy that allows fabrication of lipid membranes with designed shape by combining 3D micro-printing and replica-molding lithography with polydimethylsiloxane to create curved micrometer-sized scaffolds with virtually any geometry. The resulting supported lipid membranes are homogeneous and fluid. We demonstrate the versatility of the system by fabricating structures of interesting combinations of mean and Gaussian curvature. We study the lateral phase separation and how local curvature influences the effective diffusion coefficient. Overall, we offer a bio-compatible platform for understanding curvature-dependent cellular processes and developing programmable bio-interfaces for living cells and nanostructures.

Enzyme Related Synthesis of Lipids and Fabrication of Planar Lipid Membranes

Biological membranes are structurally and functionally complex supramolecular assemblies. The intricate complexity itself poses a significant barrier in investigating the constituents, components, and their structure-activity relationships. Scientists employ twosynthetic methods to reduce complexity: the top-down approach and the bottom-up approach. The top-down synthetic approach involves genome reduction of the living cell, leading to a collateral reduction in membrane complexity. In contrast, the bottom-upsynthetic approach like chemical and enzymatic synthesis engages assembly of non-living components to construct biomimetic membrane structures with desired complexity. The synthetic routes catalyzed by enzymes are referred to as enzymatic synthesis, while routes utilizing a combination of catalytic systems and enzymes are known as chemo-enzymatic synthesis. The lipids prepared by enzymatic or chemo-enzymatic method are essential to integrate into a membrane in a manner that makes them amenable to various analytical techniques. Planar lipid bilayer method is one such versatile technique which can be used to prepare membranes from individual lipid molecules to utilize in various analytical procedures. Two planar lipid membranes types, black lipid membrane and supported lipid membrane, are more common than other types due to their less complexity in preparation. This review discusses the enzyme related synthesis of lipids, the method of preparation and challenges involved in fabricating black lipid membrane and supported lipid membrane models.

Interaction of βL- and γ-Crystallin with Phospholipid Membrane Using Atomic Force Microscopy.

Highly concentrated lens proteins, mostly β- and γ-crystallin, are responsible for maintaining the structure and refractivity of the eye lens. However, with aging and cataract formation, β- and γ-crystallin are associated with the lens membrane or other lens proteins forming high-molecular-weight proteins, which further associate with the lens membrane, leading to light scattering and cataract development. The mechanism by which β- and γ-crystallin are associated with the lens membrane is unknown. This work aims to study the interaction of β- and γ-crystallin with the phospholipid membrane with and without cholesterol (Chol) with the overall goal of understanding the role of phospholipid and Chol in β- and γ-crystallin association with the membrane. Small unilamellar vesicles made of Chol/1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine (Chol/POPC) membranes with varying Chol content were prepared using the rapid solvent exchange method followed by probe tip sonication and then dispensed on freshly cleaved mica disk to prepare a supported lipid membrane. The βL- and γ-crystallin from the cortex of the bovine lens was used to investigate the time-dependent association of βL- and γ-crystallin with the membrane by obtaining the topographical images using atomic force microscopy. Our study showed that βL-crystallin formed semi-transmembrane defects, whereas γ-crystallin formed transmembrane defects on the phospholipid membrane. The size of semi-transmembrane defects increases significantly with incubation time when βL-crystallin interacts with the membrane. In contrast, no significant increase in transmembrane defect size was observed in the case of γ-crystallin. Our result shows that Chol inhibits the formation of membrane defects when βL- and γ-crystallin interact with the Chol/POPC membrane, where the degree of inhibition depends upon the amount of Chol content in the membrane. At a Chol/POPC mixing ratio of 0.3, membrane defects were observed when both βL- and γ-crystallin interacted with the membrane. However, at a Chol/POPC mixing ratio of 1, no association of γ-crystallin with the membrane was observed, which resulted in a defect-free membrane, and the severity of the membrane defect was decreased when βL-crystallin interacted with the membrane. The semi-transmembrane or transmembrane defects formed by the interaction of βL- and γ-crystallin on phospholipid membrane might be responsible for light scattering and cataract formation. However, Chol suppressed the formation of such defects in the membrane, likely maintaining lens membrane homeostasis and protecting against cataract formation.

Gold-Supported Lipid Membranes Formed by Redox-Triggered Vesicle Fusion on Binary Self-Assembled Monolayers: Ion-Pairing Association and Surface Hydrophilicity.

The assembly of biomimetic, planar supported lipid bilayers (SLBs) by the popular vesicle fusion method, which relies on the spontaneous adsorption and rupture of small unilamellar vesicles from aqueous solution on a solid surface, typically works with a limited range of support materials and lipid systems. We previously reported a conceptual advance in the formation of SLBs from vesicles in the gel or fluid phase using the interfacial ion-pairing association of charged phospholipid headgroups with electrochemically generated cationic ferroceniums bound to a self-assembled monolayer (SAM) chemisorbed to gold. This redox-driven approach lays down a single bilayer membrane on the SAM-modified gold surface at room temperature within minutes and is compatible with both anionic and zwitterionic phospholipids. The present work explores the effects of the surface ferrocene concentration and hydrophobicity/hydrophilicity on the formation of continuous SLBs of dialkyl phosphatidylserine, dialkyl phosphatidylglycerol, and dialkyl phosphatidylcholine using binary SAMs of ferrocenylundecanethiolate (FcC11S) and dodecanethiolate (CH3C11S) or hydroxylundecanethiolate (HOC11S) comprising different surface mole fractions of ferrocene (χFcsurf). An increase in the surface hydrophilicity and surface free energy of the FcC11S/HOC11S SAM mitigates the decrease in the attractive ion-pairing interactions resulting from a reduced χFcsurf. SLBs of ≳80% area coverage form on the FcC11S/HOC11S SAM for all the phospholipid types down to χFcsurf of at least 0.2, composition yielding a water contact angle (θW) of 44 ± 4°. By contrast, a greater number of ion-pairing interactions is required on the hydrophobic FcC11S/CH3C11S surface to drive the vesicle fusion process; bilayers or bilayer patches form at χFcsurf ≳ 0.6 (θW = 97 ± 3°). These findings will aid in tailoring the surface chemistry of redox-active modified surfaces to widen the conditions that yield supported lipid membranes.

Engineering a membrane-binding protein to trimerize and induce high membrane curvature

The annexins are a family of Ca2+-dependent peripheral membrane proteins. Several annexins are implicated in plasma membrane repair and are overexpressed in cancer cells. Annexin A4 (ANXA4) and annexin A5 (ANXA5) form trimers that induce high curvature on a membrane surface, a phenomenon deemed to accelerate membrane repair. Despite being highly homologous to ANXA4, annexin A3 (ANXA3) does not form trimers on the membrane surface. Using molecular dynamics simulations, we have reverse engineered an ANXA3-mutant to trimerize on the surface of the membrane and induce high curvature reminiscent of ANXA4. In addition, atomic force microscopy images show that, like ANXA4, the engineered protein forms crystalline arrays on a supported lipid membrane. Despite the trimer-forming and curvature-inducing properties of the engineered ANXA3, it does not accumulate near a membrane lesion in laser-punctured cells and is unable to repair the lesion. Our investigation provides insights into the factors that drive annexin-mediated membrane repair and shows that the membrane-repairing property of trimer-forming annexins also necessitates high membrane binding affinity, other than trimer formation and induction of negative membrane curvature.

Single-Molecule Imaging of Lateral Mobility and Ion Channel Activity in Lipid Bilayers using Total Internal Reflection Fluorescence (TIRF) Microscopy.

High-resolution imaging techniques have shown that many ion channels are not static, but subject to highly dynamic processes, including the transient association of pore-forming and auxiliary subunits, lateral diffusion, and clustering with other proteins. However, the relationship between lateral diffusion and function is poorly understood. To approach this problem, we describe how lateral mobility and activity of individual channels in supported lipid membranes can be monitored and correlated using total internal reflection fluorescence (TIRF) microscopy. Membranes are fabricated on an ultrathin hydrogel substrate using the droplet interface bilayer (DIB) technique. Compared to other types of model membranes, these membranes have the advantage of being mechanically robust and suitable for highly sensitive analytical techniques. This protocol measures Ca2+ ion flux through single channels by observing the fluorescence emission of a Ca2+-sensitive dye in close proximity to the membrane. In contrast to classical single-molecule tracking approaches, no fluorescent fusion proteins or labels, which can interfere with lateral movement and function in the membrane, are required. Possible changes in ion flux associated with conformational changes of the protein are only due to protein lateral motion in the membrane. Representative results are shown using the mitochondrial protein translocation channel TOM-CC and the bacterial channel OmpF. In contrast to OmpF, the gating of TOM-CC is very sensitive to molecular confinement and the nature of lateral diffusion. Hence, supported droplet-interface bilayers are a powerful tool to characterize the link between lateral diffusion and the function of ion channels.

Effect of hydrophobic and hydrophilic surfaces on the morphological and physical properties of supported lipid membranes.

The characteristics of supported lipid bilayers (SLBs) have long been studied as SLBs are commonly accepted as basic model systems of cell membranes. Atomic force microscopy (AFM) has proved to be effective in characterising physical and thermodynamic properties of SLBs, owing to its high resolution and the ability of environmental control. Through study of these properties, information such as lipid phase transitions can be investigated. Lipid phase transitions, from solid-ordered to liquid-disordered states, are well known to vary depending upon temperature, pH and ionic strength etc., and play a substantial role into events such as membrane protein mobility. Although previously studied on hydrophilic substrates, knowledge on the impact of hydrophobic substrates, such as van der Waals (vdW) crystals, on lipid phase transition is still lacking. This is particularly important for developing new bio- and electrochemical sensing technologies, where conductive vdW materials are used as sensing electrodes. It is expected that the thermodynamic properties of SLBs affect their conductive and dielectric polarisation and in turn molecular charge distribution and dielectric properties are known to strongly influence hydration layers occurring at the interface of lipids. Here, we present the study of adsorption of lipid membranes on various substrates, where morphological and physical (i.e. thermodynamic, electrical, and interfacial) properties of the membrane were investigated using advanced AFM modes as a function of temperature. We find that the transition temperature of the lipid monolayers greatly differs depending upon the chosen substrate. We then probe interfacial properties of these electrically charged interfaces. Finally, we demonstrate that substrates with different chemical-physical properties used as a lipids support have a substantial effect on the physical properties of lipid monolayers.

Beta-crystallin interacts with phospholipid membrane forming semi-transmembrane defects: A study using atomic force microscopy.

Highly concentrated lens proteins, mostly crystallins, are responsible for maintaining the structure and refractivity of the eye lens. These crystallins alter the refractivity of the lens due to self-modifications and might change the mechanical properties by interacting with the membrane. However, the interaction between a crucial lens protein, β-crystallin, with the membrane is still ambiguous. In this study, we explore the interaction of native β-crystallin extracted and purified from the cortex of the bovine lens with the 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine (POPC) membrane using atomic force microscopy (AFM). We prepared a supported lipid membrane by incubating small unilamellar vesicles (SUVs) on top of a freshly cleaved mica surface. Incubating β-crystallin solution on a defect-free membrane introduced locally modified semi-transmembrane defects. Rather than developing in multiple locations, such defects expand in the area with increased incubation time. Interestingly, the thickness mismatch between the β-crystallin-induced defects and the unaffected membrane is about half of the bilayer membrane thickness, suggesting partial removal of the distal leaflet. Force curves were obtained using force spectroscopy to access the membrane's mechanical properties. Typical membrane force curves were obtained in a defect-free region. However, the force curves obtained inside semi-transmembrane defects deviate from the characteristic membrane force curves. Membrane elasticity in the defect-free region is not significantly different from the membrane before incubating with the β-crystallin. We speculate that the defect region consists of a combination of three structural possibilities: the presence of proximal monolayer with free acyl chain, β-crystallin combined with lipids forming lipoprotein, and β-crystallin binding with lower leaflet acyl chains monolayer. Our results confirm that the interaction of β-crystallin with the POPC membrane significantly modulates membrane structure by creating semi-transmembrane defects in the supported membrane.

In Vitro Membrane Platform for the Visualization of Water Impermeability across the Liquid-Ordered Phase under Hypertonic Conditions.

Passive water penetration across the cell membrane by osmotic diffusion is essential for the homeostasis of cell volume, in addition to the protein-assisted active transportation of water. Since membrane components can regulate water permeability, controlling compositional variation during the volume regulatory process is a prerequisite for investigating the underlying mechanisms of water permeation and related membrane dynamics. However, the lack of a viable in vitro membrane platform in hypertonic solutions impedes advanced knowledge of cell volume regulation processes, especially cholesterol-enriched lipid domains called lipid rafts. By reconstituting the liquid-ordered (Lo) domain as a likeness of lipid rafts, we verified suppressed water permeation across the Lo domains, which had yet to be confirmed with experimental demonstrations despite a simulation approach. With the help of direct transfer of the Lo domains from vesicles to supported lipid membranes, the biological roles of lipid composition in suppressed water translocation were experimentally confirmed. Additionally, the improvement in membrane stability under hypertonic conditions was demonstrated based on molecular dynamics simulations.

Modulation of viscoelasticity and interfacial potential of polyelectrolyte brush by Ion-specific interactions

Polyelectrolyte brushes have drawn increasing attention because their physicochemical properties can be modulated by adjustment of the pH and ion concentration. Here, we report the controlled grafting of poly acrylic acid containing cysteine side chains onto supported lipid membranes to allow for the modulation of viscoelasticity as well as interfacial potential by ion-specific interactions, that is, with cadmium ions. Quartz crystal microbalance with dissipation indicated that the resonance frequency increased and the dissipation decreased as the cadmium concentration increased, attributed to the dehydration of brushes. Systematic variation of the molecular structure demonstrated that the coexistence of thiol and carboxyl moieties is necessary for the viscoelastic response, suggesting that these structural features, common with naturally occurring proteins, form complexes with cadmium ions. Analysis of the height fluctuation of colloidal particles by reflection interference contrast microscopy indicated that the change in the viscoelasticity of the polymer brush layer alters the curvature of the effective interfacial potential. Intriguingly, we found that modulation of the viscoelasticity and interfacial potential caused by calcium ions is weak, suggesting that the interaction is ion-specific. Polymer brushes that can alter the interfacial potential through changes in the degree of hydration opens new avenues for the design of smart, adaptable surfaces.

Systematic measurements of interleaflet friction in supported bilayers

When lipid membranes curve or are subjected to strong shear forces, the two apposed leaflets of the bilayer slide past each other. The drag that one leaflet creates on the other is quantified by the coefficient of interleaflet friction, b. Existing measurements of this coefficient range over several orders of magnitude, so we used a recently developed microfluidic technique to measure it systematically in supported lipid membranes. Fluid shear stress was used to force the top leaflet of a supported membrane to slide over the stationary lower leaflet. Here, we show that this technique yields a reproducible measurement of the friction coefficient and is sensitive enough to detect differences in friction between membranes made from saturated and unsaturated lipids. Adding cholesterol to saturated and unsaturated membranes increased interleaflet friction significantly. We also discovered that fluid shear stress can reversibly induce gel phase in supported lipid bilayers that are close to the gel-transition temperature.

An AFM Approach Applied in a Study of α-Crystallin Membrane Association: New Insights into Lens Hardening and Presbyopia Development.

The lens of the eye loses elasticity with age, while α-crystallin association with the lens membrane increases with age. It is unclear whether there is any correlation between α-crystallin association with the lens membrane and loss in lens elasticity. This research investigated α-crystallin membrane association using atomic force microscopy (AFM) for the first time to study topographical images and mechanical properties (breakthrough force and membrane area compressibility modulus (KA), as measures of elasticity) of the membrane. α-Crystallin extracted from the bovine lens cortex was incubated with a supported lipid membrane (SLM) prepared on a flat mica surface. The AFM images showed the time-dependent interaction of α-crystallin with the SLM. Force spectroscopy revealed the presence of breakthrough events in the force curves obtained in the membrane regions where no α-crystallin was associated, which suggests that the membrane’s elasticity was maintained. The force curves in the α-crystallin submerged region and the close vicinity of the α-crystallin associated region in the membrane showed no breakthrough event within the defined peak force threshold, indicating loss of membrane elasticity. Our results showed that the association of α-crystallin with the membrane deteriorates membrane elasticity, providing new insights into understanding the molecular basis of lens hardening and presbyopia.

New Insights into the Interaction of Class II Dihydroorotate Dehydrogenases with Ubiquinone in Lipid Bilayers as a Function of Lipid Composition.

The fourth enzymatic reaction in the de novo pyrimidine biosynthesis, the oxidation of dihydroorotate to orotate, is catalyzed by dihydroorotate dehydrogenase (DHODH). Enzymes belonging to the DHODH Class II are membrane-bound proteins that use ubiquinones as their electron acceptors. We have designed this study to understand the interaction of an N-terminally truncated human DHODH (HsΔ29DHODH) and the DHODH from Escherichia coli (EcDHODH) with ubiquinone (Q10) in supported lipid membranes using neutron reflectometry (NR). NR has allowed us to determine in situ, under solution conditions, how the enzymes bind to lipid membranes and to unambiguously resolve the location of Q10. Q10 is exclusively located at the center of all of the lipid bilayers investigated, and upon binding, both of the DHODHs penetrate into the hydrophobic region of the outer lipid leaflet towards the Q10. We therefore show that the interaction between the soluble enzymes and the membrane-embedded Q10 is mediated by enzyme penetration. We can also show that EcDHODH binds more efficiently to the surface of simple bilayers consisting of 1-palmitoyl, 2-oleoyl phosphatidylcholine, and tetraoleoyl cardiolipin than HsΔ29DHODH, but does not penetrate into the lipids to the same degree. Our results also highlight the importance of Q10, as well as lipid composition, on enzyme binding.

Spontaneous formation and growth kinetics of lipid nanotubules induced by passive nanoparticles.

Lipid nanotubules (LNTs) are conduits that form on the membranes of cells and organelles, and they are ubiquitous in all forms of life from archaea and bacteria to plants and mammals. The formation, shape and dynamics of these LNTs are critical for cellular functions, supporting the transport of myriad cellular cargoes as well as communication within and between cells, and they are also widely believed to be responsible for exploitation of host cells by pathogens for the spread of infection and diseases. In vitro kinetic control of LNT formation can considerably enhance the scope of utilization of these structures for disease control and therapy. Here we report a new paradigm for spontaneous lipid nanotubulation, capturing the dynamical regimes of growth, stabilization and retraction of the tubes through the binding of synthetic nanoparticles on supported lipid bilayers (SLBs). The tubulation is determined by the spontaneous binding-unbinding of nanoparticles on the LNTs. The presented methodology could be used to rectify malfunctioning cellular tubules or to prevent the pathogenic spread of diseases through inhibition of cell-to-cell nanotubule formation.

Atomic force spectroscopy with magainin 1 functionalized tips and biomimetic supported lipid membranes.

Antimicrobial peptides are molecules synthesized by living organisms as the first line of defense against bacteria, fungi, parasites, or viruses. Since their biological activity is based on destabilization of the microbial membranes, a study of direct interaction forces between antimicrobial peptides and biomimetic membranes is very important for understanding the molecular mechanisms of their action. Herein, we use atomic force spectroscopy to probe the interaction between atomic force microscopy (AFM) tips functionalized with magainin 1 and supported lipid bilayers (SLBs) mimicking electrically uncharged membranes of normal eukaryotic cells and negatively charged membranes of bacterial cells. The investigations performed on negatively charged SLBs showed that the magainin 1 functionalized AFM tips are quickly adsorbed into the SLBs when they approach, while they adhere strongly to the lipid membrane when retracted. On contrary, same investigations performed on neutral SLBs showed mechanical resistance of the lipid membrane to the tip breakthrough and negligible adhesion force at detachment.