Nanoarchitectured air-stable supported lipid bilayer incorporating sucrose–bicelle complex system

AbstractCell-membrane-mimicking supported lipid bilayers (SLBs) provide an ultrathin, self-assembled layer that forms on solid supports and can exhibit antifouling, signaling, and transport properties among various possible functions. While recent material innovations have increased the number of practically useful SLB fabrication methods, typical SLB platforms only work in aqueous environments and are prone to fluidity loss and lipid-bilayer collapse upon air exposure, which limits industrial applicability. To address this issue, herein, we developed sucrose–bicelle complex system to fabricate air-stable SLBs that were laterally mobile upon rehydration. SLBs were fabricated from bicelles in the presence of up to 40 wt% sucrose, which was verified by quartz crystal microbalance-dissipation (QCM-D) and fluorescence recovery after photobleaching (FRAP) experiments. The sucrose fraction in the system was an important factor; while 40 wt% sucrose induced lipid aggregation and defects on SLBs after the dehydration–rehydration process, 20 wt% sucrose yielded SLBs that exhibited fully recovered lateral mobility after these processes. Taken together, these findings demonstrate that sucrose–bicelle complex system can facilitate one-step fabrication of air-stable SLBs that can be useful for a wide range of biointerfacial science applications.

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Transport efficiency of membrane-anchored kinesin-1 motors depends on motor density and diffusivity

Proceedings of the National Academy of Sciences ◽

10.1073/pnas.1611398113 ◽

2016 ◽

Vol 113 (46) ◽

pp. E7185-E7193 ◽

Cited By ~ 35

Author(s):

Rahul Grover ◽

Janine Fischer ◽

Friedrich W. Schwarz ◽

Wilhelm J. Walter ◽

Petra Schwille ◽

...

Keyword(s):

Lipid Bilayer ◽

Lipid Bilayers ◽

Single Molecule ◽

Gliding Motility ◽

Supported Lipid Bilayers ◽

Transport Efficiency ◽

Collective Transport ◽

Wide Range ◽

Motor Density

In eukaryotic cells, membranous vesicles and organelles are transported by ensembles of motor proteins. These motors, such as kinesin-1, have been well characterized in vitro as single molecules or as ensembles rigidly attached to nonbiological substrates. However, the collective transport by membrane-anchored motors, that is, motors attached to a fluid lipid bilayer, is poorly understood. Here, we investigate the influence of motors’ anchorage to a lipid bilayer on the collective transport characteristics. We reconstituted “membrane-anchored” gliding motility assays using truncated kinesin-1 motors with a streptavidin-binding peptide tag that can attach to streptavidin-loaded, supported lipid bilayers. We found that the diffusing kinesin-1 motors propelled the microtubules in the presence of ATP. Notably, we found the gliding velocity of the microtubules to be strongly dependent on the number of motors and their diffusivity in the lipid bilayer. The microtubule gliding velocity increased with increasing motor density and membrane viscosity, reaching up to the stepping velocity of single motors. This finding is in contrast to conventional gliding motility assays where the density of surface-immobilized kinesin-1 motors does not influence the microtubule velocity over a wide range. We reason that the transport efficiency of membrane-anchored motors is reduced because of their slippage in the lipid bilayer, an effect that we directly observed using single-molecule fluorescence microscopy. Our results illustrate the importance of motor–cargo coupling, which potentially provides cells with an additional means of regulating the efficiency of cargo transport.

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Supported lipid bilayer repair mediated by AH peptide

Physical Chemistry Chemical Physics ◽

10.1039/c5cp06472d ◽

2016 ◽

Vol 18 (4) ◽

pp. 3040-3047 ◽

Cited By ~ 8

Author(s):

Min Chul Kim ◽

Anders Gunnarsson ◽

Seyed R. Tabaei ◽

Fredrik Höök ◽

Nam-Joon Cho

Keyword(s):

Lipid Bilayer ◽

Lipid Bilayers ◽

Silicon Oxide ◽

Supported Lipid Bilayers ◽

High Quality ◽

Supported Lipid Bilayer

High quality and complete supported lipid bilayers are formed on silicon oxide by employing an AH peptide mediated repair step.

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Transport efficiency of membrane-anchored kinesin-1 motors depends on motor density and diffusivity

10.1101/064246 ◽

2016 ◽

Cited By ~ 1

Author(s):

Rahul Grover ◽

Janine Fischer ◽

Friedrich W. Schwarz ◽

Wilhelm J. Walter ◽

Petra Schwille ◽

...

Keyword(s):

Lipid Bilayer ◽

Lipid Bilayers ◽

Gliding Motility ◽

Supported Lipid Bilayers ◽

Transport Efficiency ◽

Collective Transport ◽

Wide Range ◽

Biological Substrates ◽

Motor Density

AbstractIn eukaryotic cells, membranous vesicles and organelles are transported by ensembles of motor proteins. These motors, such as kinesin-1, have been well characterized in vitro as single molecules or as ensembles rigidly attached to non-biological substrates. However, the collective transport by membrane-anchored motors, i.e. motors attached to a fluid lipid bilayer, is poorly understood. Here, we investigate the influence of motors anchorage to a lipid bilayer on the collective transport characteristics. We reconstituted ‘membrane-anchored’ gliding motility assays using truncated kinesin-1 motors with a streptavidin-binding-peptide tag that can attach to streptavidin-loaded, supported lipid bilayers. We found that the diffusing kinesin-1 motors propelled the microtubules in presence of ATP. Notably, we found the gliding velocity of the microtubules to be strongly dependent on the number of motors and their diffusivity in the lipid bilayer. The microtubule gliding velocity increased with increasing motor density and membrane viscosity, reaching up to the stepping velocity of single-motors. This finding is in contrast to conventional gliding motility assays where the density of surface-immobilized kinesin-1 motors does not influence the microtubule velocity over a wide range. We reason, that the transport efficiency of membrane-anchored motors is reduced because of their slippage in the lipid bilayer, an effect which we directly observed using singlemolecule fluorescence microscopy. Our results illustrate the importance of the motor-cargo coupling, which potentially provides cells with an additional means of regulating the efficiency of cargo transport.

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Microcavity array supported lipid bilayer models of ganglioside – influenza hemagglutinin1 binding

Chemical Communications ◽

10.1039/d0cc04276e ◽

2020 ◽

Vol 56 (76) ◽

pp. 11251-11254 ◽

Cited By ~ 1

Author(s):

Guilherme B. Berselli ◽

Nirod Kumar Sarangi ◽

Aurélien V. Gimenez ◽

Paul V. Murphy ◽

Tia E. Keyes

Keyword(s):

Lipid Bilayer ◽

Lipid Bilayers ◽

Supported Lipid Bilayers ◽

Supported Lipid Bilayer

The binding of influenza receptor (HA1) to membranes containing different glycosphingolipid receptors was investigated at Microcavity Supported Lipid Bilayers (MSLBs).

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Influence of Brain Gangliosides on the Formation and Properties of Supported Lipid Bilayers for Binding Kinetics Studies

10.26434/chemrxiv.7505330 ◽

2018 ◽

Author(s):

Luke Jordan ◽

Nathan Wittenberg

Keyword(s):

Lipid Bilayers ◽

Binding Kinetics ◽

Supported Lipid Bilayers ◽

Dependent Manner ◽

Concentration Dependent Manner ◽

Supported Lipid Bilayer ◽

Head Groups ◽

Lipid Diffusion ◽

Kinetics Of ◽

Brain Gangliosides

This is a comprehensive study of the effects of the four major brain gangliosides (GM1, GD1b, GD1a, and GT1b) on the adsorption and rupture of phospholipid vesicles on SiO2 surfaces for the formation of supported lipid bilayer (SLB) membranes. Using quartz crystal microbalance with dissipation monitoring (QCM-D) we show that gangliosides GD1a and GT1b significantly slow the SLB formation process, whereas GM1 and GD1b have smaller effects. This is likely due to the net ganglioside charge as well as the positions of acidic sugar groups on ganglioside glycan head groups. Data is included that shows calcium can accelerate the formation of ganglioside-rich SLBs. Using fluorescence recovery after photobleaching (FRAP) we also show that the presence of gangliosides significantly reduces lipid diffusion coefficients in SLBs in a concentration-dependent manner. Finally, using QCM-D and GD1a-rich SLB membranes we measure the binding kinetics of an anti-GD1a antibody that has similarities to a monoclonal antibody that is a hallmark of a variant of Guillain-Barre syndrome.

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Lipid bilayer degradation induced by SARS-CoV-2 spike protein as revealed by neutron reflectometry

Scientific Reports ◽

10.1038/s41598-021-93996-x ◽

2021 ◽

Vol 11 (1) ◽

Author(s):

Alessandra Luchini ◽

Samantha Micciulla ◽

Giacomo Corucci ◽

Krishna Chaithanya Batchu ◽

Andreas Santamaria ◽

...

Keyword(s):

Lipid Bilayer ◽

Lipid Bilayers ◽

Membrane Lipids ◽

Lipid Extraction ◽

Specific Interaction ◽

Structural Features ◽

Supported Lipid Bilayers ◽

Fusion Event ◽

Angiotensin Converting Enzyme 2 ◽

Neutron Reflection

AbstractSARS-CoV-2 spike proteins are responsible for the membrane fusion event, which allows the virus to enter the host cell and cause infection. This process starts with the binding of the spike extramembrane domain to the angiotensin-converting enzyme 2 (ACE2), a membrane receptor highly abundant in the lungs. In this study, the extramembrane domain of SARS-CoV-2 Spike (sSpike) was injected on model membranes formed by supported lipid bilayers in presence and absence of the soluble part of receptor ACE2 (sACE2), and the structural features were studied at sub-nanometer level by neutron reflection. In all cases the presence of the protein produced a remarkable degradation of the lipid bilayer. Indeed, both for membranes from synthetic and natural lipids, a significant reduction of the surface coverage was observed. Quartz crystal microbalance measurements showed that lipid extraction starts immediately after sSpike protein injection. All measurements indicate that the presence of proteins induces the removal of membrane lipids, both in the presence and in the absence of ACE2, suggesting that sSpike molecules strongly associate with lipids, and strip them away from the bilayer, via a non-specific interaction. A cooperative effect of sACE2 and sSpike on lipid extraction was also observed.

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Lipid flip-flop and desorption from supported lipid bilayers is independent of curvature

PLoS ONE ◽

10.1371/journal.pone.0244460 ◽

2020 ◽

Vol 15 (12) ◽

pp. e0244460

Author(s):

Haoyuan Jing ◽

Yanbin Wang ◽

Parth Rakesh Desai ◽

Kumaran S. Ramamurthi ◽

Siddhartha Das

Keyword(s):

Lipid Bilayer ◽

Lipid Bilayers ◽

Md Simulations ◽

Packing Fraction ◽

Membrane Curvature ◽

Supported Lipid Bilayer ◽

Flip Flop ◽

Lipid Molecule ◽

Membrane Asymmetry ◽

Cell Shapes

Flip-flop of lipids of the lipid bilayer (LBL) constituting the plasma membrane (PM) plays a crucial role in a myriad of events ranging from cellular signaling and regulation of cell shapes to cell homeostasis, membrane asymmetry, phagocytosis, and cell apoptosis. While extensive research has been conducted to probe the lipid flip flop of planar lipid bilayers (LBLs), less is known regarding lipid flip-flop for highly curved, nanoscopic LBL systems despite the vast importance of membrane curvature in defining the morphology of cells and organelles and in maintaining a variety of cellular functions, enabling trafficking, and recruiting and localizing shape-responsive proteins. In this paper, we conduct molecular dynamics (MD) simulations to study the energetics, structure, and configuration of a lipid molecule undergoing flip-flop and desorption in a highly curved LBL, represented as a nanoparticle-supported lipid bilayer (NPSLBL) system. We compare our findings against those of a planar substrate supported lipid bilayer (PSSLBL). Our MD simulation results reveal that despite the vast differences in the curvature and other curvature-dictated properties (e.g., lipid packing fraction, difference in the number of lipids between inner and outer leaflets, etc.) between the NPSLBL and the PSSLBL, the energetics of lipid flip-flop and lipid desorption as well as the configuration of the lipid molecule undergoing lipid flip-flop are very similar for the NPSLBL and the PSSLBL. In other words, our results establish that the curvature of the LBL plays an insignificant role in lipid flip-flop and desorption.

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Monitoring supported lipid bilayers with n-type organic electrochemical transistors

Materials Horizons ◽

10.1039/d0mh00548g ◽

2020 ◽

Vol 7 (9) ◽

pp. 2348-2358 ◽

Cited By ~ 2

Author(s):

Malak Kawan ◽

Tania C. Hidalgo ◽

Weiyuan Du ◽

Anna-Maria Pappa ◽

Róisín M. Owens ◽

...

Keyword(s):

Lipid Bilayer ◽

Lipid Bilayers ◽

Supported Lipid Bilayers ◽

Accumulation Mode ◽

Electrochemical Transistor ◽

Organic Electrochemical Transistors

An n-type, accumulation mode, microscale organic electrochemical transistor monitors the activity of a pore-forming protein integrated into a lipid bilayer.

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Nanoparticle-Lipid Interaction: Job Scattering Plots to Differentiate Vesicle Aggregation from Supported Lipid Bilayer Formation

Colloids and Interfaces ◽

10.3390/colloids2040050 ◽

2018 ◽

Vol 2 (4) ◽

pp. 50 ◽

Cited By ~ 3

Author(s):

Fanny Mousseau ◽

Evdokia Oikonomou ◽

Victor Baldim ◽

Stéphane Mornet ◽

Jean-François Berret

Keyword(s):

Lipid Bilayers ◽

Lipid Vesicles ◽

Living Cells ◽

Supported Lipid Bilayers ◽

Supported Lipid Bilayer ◽

Mixed Aggregates ◽

Method Of Continuous Variation ◽

The Impact ◽

Analytical Expressions ◽

Lipid Interaction

The impact of nanomaterials on lung fluids, or on the plasma membrane of living cells, has prompted researchers to examine the interactions between nanoparticles and lipid vesicles. Recent studies have shown that nanoparticle-lipid interaction leads to a broad range of structures including supported lipid bilayers (SLB), particles adsorbed at the surface or internalized inside vesicles, and mixed aggregates. Currently, there is a need to have simple protocols that can readily evaluate the structures made from particles and vesicles. Here we apply the method of continuous variation for measuring Job scattering plots and provide analytical expressions for the scattering intensity in various scenarios. The result that emerges from the comparison between experiments and modeling is that electrostatics play a key role in the association, but it is not sufficient to induce the formation of supported lipid bilayers.

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