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

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.

Influence of Brain Gangliosides on the Formation and Properties of Supported Lipid Bilayers

10.26434/chemrxiv.7505330.v2 ◽

2019 ◽

Author(s):

Luke Jordan ◽

Megan Blauch ◽

Ashley Baxter ◽

Jennie Cawley ◽

Nathan Wittenberg

Keyword(s):

Lipid Bilayers ◽

Dependent Manner ◽

Concentration Dependent Manner ◽

Supported Lipid Bilayer ◽

Head Groups ◽

Lipid Diffusion ◽

Kinetics Of ◽

Brain Gangliosides ◽

Comprehensive Study

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.

Measuring microtubule binding kinetics of membrane-bound kinesin motors using supported lipid bilayers

STAR Protocols ◽

10.1016/j.xpro.2021.100691 ◽

2021 ◽

Vol 2 (3) ◽

pp. 100691

Author(s):

Rui Jiang ◽

William O. Hancock

Keyword(s):

Lipid Bilayers ◽

Binding Kinetics ◽

Microtubule Binding ◽

Membrane Bound ◽

Membrane-tethered mucin-like polypeptides sterically inhibit binding and slow fusion kinetics of influenza A virus

Proceedings of the National Academy of Sciences ◽

10.1073/pnas.1921962117 ◽

2020 ◽

Vol 117 (23) ◽

pp. 12643-12650 ◽

Cited By ~ 2

Author(s):

Corleone S. Delaveris ◽

Elizabeth R. Webster ◽

Steven M. Banik ◽

Steven G. Boxer ◽

Carolyn R. Bertozzi

Keyword(s):

Lipid Bilayers ◽

Influenza A ◽

Conformational Transition ◽

Molecular Model ◽

High Surface ◽

Protective Mechanism ◽

Dependent Manner ◽

Influenza A Viruses ◽

Virus Binding ◽

The mechanism(s) by which cell-tethered mucins modulate infection by influenza A viruses (IAVs) remain an open question. Mucins form both a protective barrier that can block virus binding and recruit IAVs to bind cells via the sialic acids of cell-tethered mucins. To elucidate the molecular role of mucins in flu pathogenesis, we constructed a synthetic glycocalyx to investigate membrane-tethered mucins in the context of IAV binding and fusion. We designed and synthesized lipid-tethered glycopolypeptide mimics of mucins and added them to lipid bilayers, allowing chemical control of length, glycosylation, and surface density of a model glycocalyx. We observed that the mucin mimics undergo a conformational change at high surface densities from a compact to an extended architecture. At high surface densities, asialo mucin mimics inhibited IAV binding to underlying glycolipid receptors, and this density correlated to the mucin mimic’s conformational transition. Using a single virus fusion assay, we observed that while fusion of virions bound to vesicles coated with sialylated mucin mimics was possible, the kinetics of fusion was slowed in a mucin density-dependent manner. These data provide a molecular model for a protective mechanism by mucins in IAV infection, and therefore this synthetic glycocalyx provides a useful reductionist model for studying the complex interface of host–pathogen interactions.

Leukotriene B4 stimulates c-fos and c-jun gene transcription and AP-1 binding activity in human monocytes

Biochemical Journal ◽

10.1042/bj2820625 ◽

1992 ◽

Vol 282 (3) ◽

pp. 625-629 ◽

Cited By ~ 39

Author(s):

J Staňková ◽

M Rola-Pleszczynski

Keyword(s):

Human Peripheral Blood ◽

Leukotriene B4 ◽

Binding Activity ◽

Dependent Manner ◽

Blood Monocytes ◽

Concentration Dependent Manner ◽

Nuclear Factors ◽

Oncogene Products ◽

We have examined the effect of leukotriene B4 (LTB4), a potent lipid proinflammatory mediator, on the expression of the proto-oncogenes c-jun and c-fos. In addition, we looked at the modulation of nuclear factors binding specifically to the AP-1 element after LTB4 stimulation. LTB4 increased the expression of the c-fos gene in a time- and concentration-dependent manner. The c-jun mRNA, which is constitutively expressed in human peripheral-blood monocytes at relatively high levels, was also slightly augmented by LTB4, although to a much lower extent than c-fos. The kinetics of expression of the two genes were also slightly different, with c-fos mRNA reaching a peak at 15 min after stimulation and c-jun at 30 min. Both messages rapidly declined thereafter. Stability of the c-fos and c-jun mRNA was not affected by LTB4, as assessed after actinomycin D treatment. Nuclear transcription studies in vitro showed that LTB4 increased the transcription of the c-fos gene 7-fold and the c-jun gene 1.4-fold. Resting monocytes contained nuclear factors binding to the AP-1 element, but stimulation of monocytes with LTB4 induced greater AP-1-binding activity of nuclear proteins. These results indicate that LTB4 may regulate the production of different cytokines by modulating the yield and/or the function of transcription factors such as AP-1-binding proto-oncogene products.

On the Kinetics of Adsorption and Two-Dimensional Self-Assembly of Annexin A5 on Supported Lipid Bilayers

Biophysical Journal ◽

10.1529/biophysj.105.064337 ◽

2005 ◽

Vol 89 (5) ◽

pp. 3372-3385 ◽

Cited By ~ 96

Author(s):

Ralf P. Richter ◽

Joséphine Lai Kee Him ◽

Béatrice Tessier ◽

Céline Tessier ◽

Alain R. Brisson

Keyword(s):

Lipid Bilayers ◽

Self Assembly ◽

Annexin A5 ◽

Two Dimensional ◽

Kinetics Of Adsorption ◽

Membrane-Tethered Mucin-like Polypeptides Sterically Inhibit Binding and Slow Fusion Kinetics of Influenza A Virus

10.26434/chemrxiv.11312567 ◽

2019 ◽

Cited By ~ 1

Author(s):

Corleone Delaveris ◽

Elizabeth Webster ◽

Steven Banik ◽

Steven Boxer ◽

Carolyn Bertozzi

Keyword(s):

Lipid Bilayers ◽

Influenza A ◽

Conformational Transition ◽

Molecular Model ◽

High Surface ◽

Protective Mechanism ◽

Dependent Manner ◽

Influenza A Viruses ◽

Virus Binding ◽

<div> <div> <div> <p>The mechanism(s) by which cell-tethered mucins modulate infection by Influenza A viruses (IAVs) remains an open question. Mucins form both a protective barrier that can block virus binding and recruit IAVs to bind cells via the sialic acids of cell-tethered mucins. To elucidate the molecular role of mucins in flu pathogenesis, we constructed a synthetic glycocalyx to investigate membrane-tethered mucins in the context of IAV binding and fusion. We designed and synthesized lipid-tethered glycopolypeptide mimics of mucins and added them to lipid bilayers, allowing chemical control of length, glycosylation, and surface density of a model glycocalyx. We observed that the mucin mimics undergo a conformational change at high surface densities from a compact to an extended architecture. At high surface densities asialo mucin mimics inhibited IAV binding to underlying glycolipid receptors and this density correlated to the mucin mimic’s conformational transition. Using a single virus fusion assay, we observed that while fusion of virions bound to vesicles coated with sialylated mucin mimics was possible, the kinetics of fusion were slowed in a mucin density-dependent manner. These data provide a molecular model for a protective mechanism by mucins in IAV infection, and therefore this synthetic glycocalyx provides a useful reductionist model for studying the complex interface of host-pathogen interactions. </p> </div> </div> </div>

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 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.

cADP-ribose activates reconstituted ryanodine receptors from coronary arterial smooth muscle

AJP Heart and Circulatory Physiology ◽

10.1152/ajpheart.2001.280.1.h208 ◽

2001 ◽

Vol 280 (1) ◽

pp. H208-H215 ◽

Cited By ~ 41

Author(s):

Pin-Lan Li ◽

Wang-Xian Tang ◽

Hector H. Valdivia ◽

Ai-Ping Zou ◽

William B. Campbell

Keyword(s):

Smooth Muscle ◽

Lipid Bilayers ◽

Ryanodine Receptors ◽

Fold Increase ◽

Ruthenium Red ◽

Dependent Manner ◽

High Concentration ◽

Open Probability ◽

Arterial Smooth Muscle ◽

Concentration Dependent Manner

The present study was designed to test the hypothesis that cADP-ribose (cADPR) increases Ca2+release through activation of ryanodine receptors (RYR) on the sarcoplasmic reticulum (SR) in coronary arterial smooth muscle cells (CASMCs). We reconstituted RYR from the SR of CASMCs into planar lipid bilayers and examined the effect of cADPR on the activity of these Ca2+ release channels. In a symmetrical cesium methanesulfonate configuration, a 245 pS Cs+ current was recorded. This current was characterized by the formation of a subconductance and increase in the open probability (NPo) of the channels in the presence of ryanodine (0.01–1 μM) and imperatoxin A (100 nM). A high concentration of ryanodine (50 μM) and ruthenium red (40–80 μM) substantially inhibited the activity of RYR/Ca2+ release channels. Caffeine (0.5–5 mM) markedly increased the NPo of these Ca2+release channels of the SR, but d- myo-inositol 1,4,5-trisphospate and heparin were without effect. Cyclic ADPR significantly increased the NPo of these Ca2+release channels of SR in a concentration-dependent manner. Addition of cADPR (0.01 μM) into the cis bath solution produced a 2.9-fold increase in the NPo of these RYR/Ca2+release channels. An eightfold increase in the NPo of the RYR/Ca2+ release channels (0.0056 ± 0.001 vs. 0.048 ± 0.017) was observed at a concentration of cADPR of 1 μM. The effect of cADPR was completely abolished by ryanodine (50 μM). In the presence of cADPR, Ca2+-induced activation of these channels was markedly enhanced. These results provide evidence that cADPR activates RYR/Ca2+ release channels on the SR of CASMCs. It is concluded that cADPR stimulates Ca2+ release through the activation of RYRs on the SR of these smooth mucle cells.