Amphipathic DNA Origami Nanoparticles to Scaffold and Deform Lipid Membrane Vesicles

2015 ◽  
Vol 54 (22) ◽  
pp. 6501-6505 ◽  
Author(s):  
Aleksander Czogalla ◽  
Dominik J. Kauert ◽  
Henri G. Franquelim ◽  
Veselina Uzunova ◽  
Yixin Zhang ◽  
...  
Keyword(s):  
Lipid Membrane ◽  
Membrane Vesicles ◽  
Dna Origami

Amphipathic DNA Origami Nanoparticles to Scaffold and Deform Lipid Membrane Vesicles

2015 ◽  
Vol 127 (22) ◽  
pp. 6601-6605 ◽  
Author(s):  
Aleksander Czogalla ◽  
Dominik J. Kauert ◽  
Henri G. Franquelim ◽  
Veselina Uzunova ◽  
Yixin Zhang ◽  
...  
Keyword(s):  
Lipid Membrane ◽  
Membrane Vesicles ◽  
Dna Origami

Electrophysiological Characterization of Transport Across Outer Membrane Channels from Gram-Negative Bacteria in Their Native Environment

2019 ◽  
Author(s):  
Jiajun Wang ◽  
Rémi Terrasse ◽  
Jayesh Arun Bafna ◽  
Lorraine Benier ◽  
Mathias Winterhalter
Keyword(s):  
Outer Membrane ◽  
Lipid Membrane ◽  
Membrane Vesicles ◽  
Outer Membrane Vesicles ◽  
Multi Drug Resistance ◽  
Gram Negative Bacteria ◽  
Membrane Channels ◽  
Gram Negative ◽  
Electrophysiological Characterization

Multi-drug resistance in Gram-negative bacteria is often associated with low permeability of the outer membrane. To investigate the role of membrane channels in the uptake of antibiotics, we extract, purify and reconstitute them into artificial planar membranes. To avoid this time-consuming procedure, here we show a robust approach using fusion of native outer membrane vesicles (OMV) into planar lipid bilayer which moreover allows also to some extend the characterization of membrane protein channels in their native environment. Two major membrane channels from <i>Escherichia coli</i>, OmpF and OmpC, were overexpressed from the host and the corresponding OMVs were collected. Each OMV fusion revealed surprisingly single or only few channel activities. The asymmetry of the OMV´s translates after fusion into the lipid membrane with the LPS dominantly present at the side of OMV addition. Compared to conventional reconstitution methods, the channels fused from OMVs containing LPS have similar conductance but a much broader distribution. The addition of Enrofloxacin on the LPS side yields somewhat higher association (<i>k<sub>on</sub></i>) and lower dissociation (<i>k<sub>off</sub></i>) rates compared to LPS-free reconstitution. We conclude that using outer membrane vesicles is a fast and easy approach for functional and structural studies of membrane channels in the native membrane.

scholarly journals On the Choice of the Extracellular Vesicles for Therapeutic Purposes

2019 ◽  
Vol 20 (2) ◽  
pp. 236 ◽  
Author(s):  
Claudia Campanella ◽  
Celeste Caruso Bavisotto ◽  
Mariantonia Logozzi ◽  
Antonella Marino Gammazza ◽  
Davide Mizzoni ◽  
...  
Keyword(s):  
Regenerative Medicine ◽  
Extracellular Vesicles ◽  
Lipid Membrane ◽  
Cell Communication ◽  
Membrane Vesicles ◽  
Free Form ◽  
Target Cells ◽  
Disease Biomarkers ◽  
Cellular Source ◽  
Therapeutic Molecules

Extracellular vesicles (EVs) are lipid membrane vesicles released by all human cells and are widely recognized to be involved in many cellular processes, both in physiological and pathological conditions. They are mediators of cell-cell communication, at both paracrine and systemic levels, and therefore they are active players in cell differentiation, tissue homeostasis, and organ remodeling. Due to their ability to serve as a cargo for proteins, lipids, and nucleic acids, which often reflects the cellular source, they should be considered the future of the natural nanodelivery of bio-compounds. To date, natural nanovesicles, such as exosomes, have been shown to represent a source of disease biomarkers and have high potential benefits in regenerative medicine. Indeed, they deliver both chemical and bio-molecules in a way that within exosomes drugs are more effective that in their exosome-free form. Thus, to date, we know that exosomes are shuttle disease biomarkers and probably the most effective way to deliver therapeutic molecules within target cells. However, we do not know exactly which exosomes may be used in therapy in avoiding side effects as well. In regenerative medicine, it will be ideal to use autologous exosomes, but it seems not ideal to use plasma-derived exosomes, as they may contain potentially dangerous molecules. Here, we want to present and discuss a contradictory relatively unmet issue that is the lack of a general agreement on the choice for the source of extracellular vesicles for therapeutic use.

scholarly journals Modulation of host lipid membrane elasticity and dipole potential induced by bacterial membrane vesicles

2019 ◽  
Author(s):  
Ashutosh Prince ◽  
Anuj Tiwari ◽  
Titas Mandal ◽  
Kuldeep Sharma ◽  
Nikhil Kanik ◽  
...  
Keyword(s):  
Physicochemical Properties ◽  
Lipid Membranes ◽  
Lipid Membrane ◽  
Membrane Vesicles ◽  
Bacterial Membrane ◽  
Dipole Potential ◽  
Long Distance ◽  
Biologically Relevant ◽  
Host Membrane ◽  
Distance Delivery

AbstractBacterial membrane vesicles (MVs) facilitate long-distance delivery of virulence factors crucial for pathogenicity. The entry and trafficking mechanisms of virulence factors inside host cells is recently emerging, however, if bacterial MVs modulate the physicochemical properties of the host lipid membrane remains unknown. Here we reconstitute the interaction of bacterial MV with host cell lipid membranes and quantitatively show that bacterial MV interaction increases the fluidity, dipole potential and elasticity of a biologically relevant multi-component host membrane. The presence of cylindrical lipids such as phosphatidylcholine and phosphatidylinositol and a moderate acyl chain length of C16 helps the MV interaction. While significant binding of bacterial MVs to the raft-like lipid membranes with phase separated regions of the membrane was observed, however, MVs have a preference for binding to the liquid disordered regions of the membrane. Further, the elevated levels of cholesterol tend to hinder the interaction of bacterial MVs. We further quantify the change in excess Gibbs free energy of mixing of bacterial MVs with host lipid membranes driving the modulation of host membrane parameters. The findings may have significant implications on the regulation of host machineries by pathogen through manipulation of host membrane properties.Significance StatementBacterial membrane vesicles (MVs) act as the long-distance delivery tools for virulence factor and thus, directly implicated in host-pathogen interactions and pathogenicity. While the mechanisms of virulence transfer is only recently emerging, however, the interaction of MVs the host cell membrane remains largely unexplored. Whether the MVs interaction can locally modulate the host lipid membrane physicochemical properties (such as fluidity, dipole potential and elasticity) remains unknown. Here, we quantitatively investigate the lipid specificity of E. Coli MV interaction and this results in increase in the fluidity, dipole potential and in-plane elasticity of a biologically relevant multi-component host membrane. The findings could be important for numerous cell-signaling processes as well as downstream events involving membrane-membrane fusion during process of phagosome maturation.

scholarly journals Extracellular Vesicles in Tumors: A Potential Mediator of Bone Metastasis

2021 ◽  
Vol 9 ◽  
Author(s):  
Shenglong Li ◽  
Wei Wang
Keyword(s):  
Bone Metastasis ◽  
Extracellular Vesicles ◽  
Lipid Membrane ◽  
Bone Cells ◽  
Membrane Vesicles ◽  
Metastatic Tumor ◽  
Cell Types ◽  
Biological Phenomena ◽  
Breast And Prostate Cancer ◽  
Metastatic Sites

As one of the most common metastatic sites, bone has a unique microenvironment for the growth and prosperity of metastatic tumor cells. Bone metastasis is a common complication for tumor patients and accounts for 15–20% of systemic metastasis, which is only secondary to lung and liver metastasis. Cancers prone to bone metastasis include lung, breast, and prostate cancer. Extracellular vesicles (EVs) are lipid membrane vesicles released from different cell types. It is clear that EVs are associated with multiple biological phenomena and are crucial for intracellular communication by transporting intracellular substances. Recent studies have implicated EVs in the development of cancer. However, the potential roles of EVs in the pathological exchange of bone cells between tumors and the bone microenvironment remain an emerging area. This review is focused on the role of tumor-derived EVs in bone metastasis and possible regulatory mechanisms.

scholarly journals Polarity sensitive probes for super resolution STED microscopy

2017 ◽  
Author(s):  
E Sezgin ◽  
F Schneider ◽  
V Zilles ◽  
E Garcia ◽  
D Waithe ◽  
...  
Keyword(s):  
Plasma Membrane ◽  
Lipid Membrane ◽  
Membrane Lipids ◽  
Super Resolution ◽  
Membrane Vesicles ◽  
Molecular Organization ◽  
Live Cells ◽  
Sted Microscopy ◽  
Two Photon Microscopy ◽  
Polarity Sensitive

AbstractThe lateral organization of molecules in the cellular plasma membrane plays an important role in cellular signaling. A critical parameter for membrane molecular organization is how the membrane lipids are packed (or ordered). Polarity sensitive dyes are powerful tools to characterize such lipid membrane order, employing for example confocal and two-photon microscopy. The investigation of potential lipid nanodomains, however, requires the use of super resolution microscopy. Here, we test the performance of the polarity sensitive membrane dyes Di-4-ANEPPDHQ, Di-4-AN(F)EPPTEA and NR12S in super resolution STED microscopy. Measurements on cell-derived membrane vesicles, in the plasma membrane of live cells, and on single virus particles show the high potential of these dyes for probing nanoscale membrane heterogeneity.

Sealable Large Pore by DNA Origami on Lipid Membrane

2020 ◽  
Author(s):  
Shoji Iwabuchi ◽  
Ibuki Kawamata ◽  
Satoshi Murata ◽  
Shin-ichiro Nomura
Keyword(s):  
Cross Section ◽  
Lipid Membrane ◽  
Dna Origami ◽  
Single Strand ◽  
Vesicle Membrane ◽  
Giant Vesicle ◽  
Large Pore ◽  
And Function ◽  
Fluorescent Molecules

Here, we report on the design and function of a membrane nanopore through a DNA origami square tube with a cross-section of 100 nm2 . When the nanopore is added onto the giant vesicle membrane, the permeation of hydrophilic fluorescent molecules was observed. It can be sealed by the existence of the four specific single strand DNAs. A controllable artificial nanopore should help to communicate the vesicle components with their environment

Sealable Large Pore by DNA Origami on Lipid Membrane

2020 ◽  
Author(s):  
Shoji Iwabuchi ◽  
Ibuki Kawamata ◽  
Satoshi Murata ◽  
Shin-ichiro Nomura
Keyword(s):  
Cross Section ◽  
Lipid Membrane ◽  
Dna Origami ◽  
Single Strand ◽  
Vesicle Membrane ◽  
Giant Vesicle ◽  
Large Pore ◽  
And Function ◽  
Fluorescent Molecules

Here, we report on the design and function of a membrane nanopore through a DNA origami square tube with a cross-section of 100 nm2 . When the nanopore is added onto the giant vesicle membrane, the permeation of hydrophilic fluorescent molecules was observed. It can be sealed by the existence of the four specific single strand DNAs. A controllable artificial nanopore should help to communicate the vesicle components with their environment

scholarly journals Shear-driven circulation patterns in lipid membrane vesicles

2012 ◽  
Vol 705 ◽  
pp. 165-175 ◽  
Author(s):  
Francis G. Woodhouse ◽  
Raymond E. Goldstein
Keyword(s):  
Lipid Membrane ◽  
Membrane Vesicles ◽  
Lipid Vesicle ◽  
Simple Shear Flow ◽  
Immiscible Fluid ◽  
Marked Contrast ◽  
Membrane Viscosity ◽  
Systematic Expansion ◽  
Circulation Patterns ◽  
Dipole Vortex

AbstractRecent experiments have shown that when a near-hemispherical lipid vesicle attached to a solid surface is subjected to a simple shear flow it exhibits a pattern of membrane circulation much like a dipole vortex. This is in marked contrast to the toroidal circulation that would occur in the related problem of a drop of immiscible fluid attached to a surface and subjected to shear. This profound difference in flow patterns arises from the lateral incompressibility of the membrane, which restricts the observable flows to those in which the velocity field in the membrane is two-dimensionally divergence free. Here we study these circulation patterns within the simplest model of membrane fluid dynamics. A systematic expansion of the flow field based on Papkovich–Neuber potentials is developed for general viscosity ratios between the membrane and the surrounding fluids. Comparison with experimental results (Vézy, Massiera & Viallat, Soft Matt., vol. 3, 2007, pp. 844–851) is made, and it is shown how such studies could allow measurements of the membrane viscosity. Issues of symmetry-breaking and pattern selection are discussed.

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