Supported Lipid Bilayers of Escherichia coli Extracted Lipids and Their Calcium Dependence

Kakimoto, Yasuhiro; Tero, Ryugo

doi:10.3389/fmats.2018.00048

Cited by 8 publications

(13 citation statements)

References 54 publications

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“…Asymmetric bilayer formation − and LPS membrane models containing O-antigen − are recent developments in the field. Outer membrane models have been used to study structural and organizational roles of divalent cations such as magnesium and calcium, , the protective role of LPS against cationic antimicrobial peptides, antibiotic lipid interactions, , molecular interactions between bacterial phospholipids and LPS as it pertains to lipid packing and stabilization, , and the role of membrane lipids in protein stability and function . LPS comprises approximately 75% of the surface area of the OM and plays a role in mediating interactions with antimicrobial compounds .…”

Section: Resultsmentioning

confidence: 99%

Clinically Relevant Bacterial Outer Membrane Models for Antibiotic Screening Applications

et al. 2021

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Antibiotic resistance is a growing global health concern that has been increasing in prevalence over the past few decades. In Gram-negative bacteria, the outer membrane is an additional barrier through which antibiotics must traverse to kill the bacterium. In addition, outer membrane features and properties, like membrane surface charge, lipopolysaccharide (LPS) length, and membrane porins, can be altered in response to antibiotics and therefore, further mediate resistance. Model membranes have been used to mimic bacterial membranes to study antibiotic-induced membrane changes but often lack the compositional complexity of the actual outer membrane. Here, we developed a surface-supported membrane platform using outer membrane vesicles (OMVs) from clinically relevant Gram-negative bacteria and use it to characterize membrane biophysical properties and investigate its interaction with antibacterial compounds. We demonstrate that this platform maintains critical features of outer membranes, like fluidity, while retaining complex membrane components, like OMPs and LPS, which are central to membrane-mediated antibiotic resistance. This platform offers a non-pathogenic, cell-free surface to study such phenomena that is compatible with advanced microscopy and surface characterization tools like quartz crystal microbalance. We confirm these OMV bilayers recapitulate membrane interactions (or lack thereof) with the antibiotic compounds polymyxin B, bacitracin, and vancomycin, validating their use as representative models for the bacterial surface. By forming OMV bilayers from different strains, we envision that this platform could be used to investigate underlying biophysical differences in outer membranes leading to resistance, to screen and identify membrane-active antibiotics, or for the development of phage technologies targeting a particular membrane surface component.

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Section: Resultsmentioning

confidence: 99%

Clinically Relevant Bacterial Outer Membrane Models for Antibiotic Screening Applications

et al. 2021

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“…Divalent cation attenuated the electrostatic repulsion between DOPG (−) and the dissociated hydroxyl group (−O − ) on the SiO 2 /Si substrate, and furthermore bridged these monovalent anions. 44,45) A planar SLB is formed through adsorption, fusion, rupture, and spreading of vesicles. During these processes, a part of lipids in the adsorbed vesicles desorbs from the substrates as reported in previous QCM studies.…”

Section: Resultsmentioning

confidence: 99%

Substrate-induced electrostatic potential varies composition of supported lipid bilayer containing anionic lipid

Tero

Kobayashi

2022

Jpn. J. Appl. Phys.

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Supported lipid bilayers (SLBs) are artificial lipid bilayers at solid-liquid interfaces applied as cell membrane model systems. An advantage of the artificial system is that the lipid composition can be controlled arbitrarily. On the other hand, the SLB formation process and its efficiency are affected by the properties of the solid substrate surface. In this study, we investigated the effect of the electrostatic interaction between the negatively charged SiO2/Si substrate surface and the lipid bilayer membrane on the composition of binary SLBs comprising anionic and neutral lipids. The phase transition temperature and the area fraction of lipid domains of SLB were evaluated by fluorescence microscopy and the fluorescence recovery after photobleaching. The neutral lipid was preferably included in SLB, but the anionic lipid ratio increased with Ca2+ concentration during the SLB formation. The lipid composition in SLB can be controlled by modulating the substrate-induced electrostatic potential.

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“…Bacterial lipid extracts can be more challenging due to high quantities of non-bilayer forming lipids such as PE, and negatively charged lipids that repel the surface and require addition of e.g. Ca2+ ions, [133] but SLB formation is routinely achievable, with E.coli TLE and PLE most commonly used. [134,135] AFM analysis of pore-forming proteins and pore-forming toxins have been conducted on such bacterial lipid extracts [136,137] but the use of lipid extracts SLBs in AFM studies of peptide-induced membrane disruption has been limited, despite being used by other techniques (see e.g.…”

Section: Improved Model Systemsmentioning

confidence: 99%

Atomic force microscopy to elucidate how peptides disrupt membranes

Hammond

Ryadnov

Hoogenboom

2021

Biochimica et Biophysica Acta (BBA) - Biomembranes

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Atomic force microscopy is an increasingly attractive tool to study how peptides disrupt membranes.Often performed on reconstituted lipid bilayers, it provides access to time and length scales that allow dynamic investigations with nanometre resolution. Over the last decade, AFM studies have enabled visualisation of membrane disruption mechanisms by antimicrobial or host defence peptides, including peptides that target malignant cells and biofilms. Moreover, the emergence of high-speed modalities of the technique broadens the scope of investigations to antimicrobial kinetics as well as the imaging of peptide action on live cells in real time. This review describes how methodological advances in AFM facilitate new insights into membrane disruption mechanisms.

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Supported Lipid Bilayers of Escherichia coli Extracted Lipids and Their Calcium Dependence

Cited by 8 publications

References 54 publications

Clinically Relevant Bacterial Outer Membrane Models for Antibiotic Screening Applications

Clinically Relevant Bacterial Outer Membrane Models for Antibiotic Screening Applications

Substrate-induced electrostatic potential varies composition of supported lipid bilayer containing anionic lipid

Atomic force microscopy to elucidate how peptides disrupt membranes

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