Calcium Dependent Reversible Aggregation of Escherichia coli Biomimicking Vesicles Enables Formation of Supported Vesicle Layers on Silicon Dioxide

Duša, Filip; Chen, Wen; Witos, Joanna; Wiedmer, Susanne Κ.

doi:10.3389/fmats.2019.00023

Cited by 6 publications

(10 citation statements)

References 30 publications

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“…Recently, it was shown that CaCl 2 induced an apparent size increase in vesicle suspensions made from E. coli extracts; however, the size increase was mainly due to vesicle aggregation and not due to vesicle fusion, and this vesicle aggregation was reversible upon diluting with Ca 2+ -free buffer. 24 Here, CaCl 2 was used as the fusion promotor, and several concentrations were tested for optimizing the deposition with NR and QCM-D connected in series on the NR beamline. SLB formation via vesicle fusion is facilitated by using lipids in the fluid phase.…”

Section: Resultsmentioning

confidence: 99%

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Formation and Characterization of Supported Lipid Bilayers Composed of Phosphatidylethanolamine and Phosphatidylglycerol by Vesicle Fusion, a Simple but Relevant Model for Bacterial Membranes

2019

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Supported lipid bilayers (SLBs) are simple and robust biomimics with controlled lipid composition that are widely used as models of both mammalian and bacterial membranes. However, the lipids typically used for SLB formation poorly resemble those of bacterial cell membranes due to the lack of available protocols to form SLBs using mixtures of lipids relevant for bacteria such as phosphatidylethanolamine (PE) and phosphatidylglycerol (PG). Although a few reports have been published recently on the formation of SLBs from Escherichia coli lipid extracts, a detailed understanding of these systems is challenging due to the complexity of the lipid composition in such natural extracts. Here, we present for the first time a simple and reliable protocol optimized to form high-quality SLBs using mixtures of PE and PG at compositions relevant for Gram-negative membranes. We show using neutron reflection and quartz microbalance not only that Ca 2+ ions and temperature are key parameters for successful bilayer deposition but also that mass transfer to the surface is a limiting factor. Continuous flow of the lipid suspension is thus crucial for obtaining full SLB coverage. We furthermore characterize the resulting bilayers and report structural parameters, for the first time for PE and PG mixtures, which are in good agreement with those reported earlier for pure POPE vesicles. With this protocol in place, more suitable and reproducible studies can be conducted to understand biomolecular processes occurring at cell membranes, for example, for testing specificities and to unravel the mechanism of interaction of antimicrobial peptides.

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Section: Resultsmentioning

confidence: 99%

“…Prolonged vesicle incubation was also shown to lead to irreversible vesicle binding of E. coli extracts in the presence of CaCl 2 . 24…”

Section: Resultsmentioning

confidence: 99%

Formation and Characterization of Supported Lipid Bilayers Composed of Phosphatidylethanolamine and Phosphatidylglycerol by Vesicle Fusion, a Simple but Relevant Model for Bacterial Membranes

2019

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“…The preparations of egg PC and E. coli liposomes have been described previously in detail [12,28]. To prepare yeast liposomes, 400 µL of yeast lipid extract in chloroform (25 mg/mL) was added to a test tube.…”

Section: Liposome Preparationmentioning

confidence: 99%

“…The immobilization of egg PC and E. coli liposomes was conducted according to our previous research [12,28].…”

Section: Immobilization Of Liposomesmentioning

confidence: 99%

“…Recently, we successfully immobilized liposomes with a composition very similar to the cell membrane of Escherichia coli (E. coli) [12], when a total lipid extract from E. coli was used for the membrane preparation. The liposomes were immobilized onto silicon dioxide (SiO2)-coated sensors using sodium N-(2hydroxyethyl)piperazine-N'-2-ethane sulfonate (Na-HEPES) buffer containing calcium chloride (Ca-HEPES).…”

Section: Introductionmentioning

confidence: 99%

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Immobilization of natural lipid biomembranes and their interactions with choline carboxylates. A nanoplasmonic sensing study

Duša

Chen

Witos

et al. 2020

Biochimica et Biophysica Acta (BBA) - Biomembranes

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The cell membrane is mainly composed of lipid bilayers with inserted proteins and carbohydrates. Lipid bilayers made of purified or synthetic lipids are widely used for estimating the effect of target compounds on cell membranes. However, the composition of such biomimetic membranes is much simpler than the composition of biological membranes. Interactions between compounds and simple composition biomimetic membranes might not demonstrate the effect of target compounds as precisely as membranes with compositions close to real organisms. Therefore, the aim of our study is to construct biomimetic membrane closely mimicking the state of natural membranes. Liposomes were prepared from lipids extracted from L-α-phosphatidylcholine, Escherichia coli, yeast (Saccharomyces cerevisiae) and bovine liver cells through agitation and sonication. They were immobilized onto SiO2 sensor surfaces using N-(2-hydroxyethyl)piperazine-N'-2-ethanesulfonic acid buffer with calcium chloride. The biomimetic membranes were successfully immobilized onto the SiO2 sensor surface and detected by nanoplasmonic sensing. The immobilized membranes were exposed to choline carboxylates. The membrane disruption effect was, as expected, more pronounced with increasing carbohydrate chain length of the carboxylates. The results correlated with the toxicity values determined using Vibrio fischeri bacteria. The yeast extracted lipid membranes had the strongest response to introduction of choline laurate while the bovine liver lipid extracted liposomes were the most sensitive towards the shorter choline carboxylates. This implies that the composition of the cell membrane plays a crucial role upon interaction with choline carboxylates, and underlines the necessity of testing membrane systems of different origin to obtain an overall image of such interactions.

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Design and use of model membranes to study biomolecular interactions using complementary surface-sensitive techniques

Clifton

Campbell

Sebastiani

et al. 2020

Advances in Colloid and Interface Science

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Highlights• Model biomembranes at surfaces are useful to study biomolecular interactions • The model membrane complexity can be adapted from one to multiple components • Implementations range from lipid monolayers to a range of supported bilayers • Complementary surface-sensitive techniques allow studying model membranes • Information accessible is reviewed via the use of the peptide Melittin

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Calcium Dependent Reversible Aggregation of Escherichia coli Biomimicking Vesicles Enables Formation of Supported Vesicle Layers on Silicon Dioxide

Cited by 6 publications

References 30 publications

Formation and Characterization of Supported Lipid Bilayers Composed of Phosphatidylethanolamine and Phosphatidylglycerol by Vesicle Fusion, a Simple but Relevant Model for Bacterial Membranes

Formation and Characterization of Supported Lipid Bilayers Composed of Phosphatidylethanolamine and Phosphatidylglycerol by Vesicle Fusion, a Simple but Relevant Model for Bacterial Membranes

Immobilization of natural lipid biomembranes and their interactions with choline carboxylates. A nanoplasmonic sensing study

Design and use of model membranes to study biomolecular interactions using complementary surface-sensitive techniques

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