Fabrication of highly insulating tethered bilayer lipid membrane using yeast cell membrane fractions for measuring ion channel activity

Jadhav, Sachin R.; Sui, Dexin; Garavito, R. Michael; Worden, R. Mark

doi:10.1016/j.jcis.2008.02.064

Cited by 38 publications

(32 citation statements)

References 37 publications

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“…This flexibility enables the determination of binding saturation values that are more representative of biological systems. In addition to model membranes, sensing platforms can consist of cell-derived membranes that contain a natural assortment of components including lipids, proteins, and glycosaminoglycans [77][78][79]. While model membranes represent a general platform to study membrane association processes driven by amphiphilic interactions, cell-derived membranes contain receptors which can mediate specific binding of proteins.…”

Section: Engineering Strategy To Mimic Biological Membranesmentioning

confidence: 99%

Model Membrane Platforms for Biomedicine: Case Study on Antiviral Drug Development

Jackman

Cho

2012

Biointerphases

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As one of the most important interfaces in cellular systems, biological membranes have essential functions in many activities such as cellular protection and signaling. Beyond their direct functions, they also serve as scaffolds to support the association of proteins involved in structural support, adhesion, and transport. Unfortunately, biological processes sometimes malfunction and require therapeutic intervention. For those processes which occur within or upon membranes, it is oftentimes difficult to study the mechanism in a biologically relevant, membranous environment. Therefore, the identification of direct therapeutic targets is challenging. In order to overcome this barrier, engineering strategies offer a new approach to interrogate biological activities at membrane interfaces by analyzing them through the principles of the interfacial sciences. Since membranes are complex biological interfaces, the development of simplified model systems which mimic important properties of membranes can enable fundamental characterization of interaction parameters for such processes. We have selected the hepatitis C virus (HCV) as a model viral pathogen to demonstrate how model membrane platforms can aid antiviral drug discovery and development. Responsible for generating the genomic diversity that makes treating HCV infection so difficult, viral replication represents an ideal step in the virus life cycle for therapeutic intervention. To target HCV genome replication, the interaction of viral proteins with model membrane platforms has served as a useful strategy for target identification and characterization. In this review article, we demonstrate how engineering approaches have led to the discovery of a new functional activity encoded within the HCV nonstructural 5A protein. Specifically, its N-terminal amphipathic, α-helix (AH) can rupture lipid vesicles in a size-dependent manner. While this activity has a number of exciting biotechnology and biomedical applications, arguably the most promising one is in antiviral medicine. Based on the similarities between lipid vesicles and the lipid envelopes of virus particles, experimental findings from model membrane platforms led to the prediction that a range of medically important viruses might be susceptible to rupturing treatment with synthetic AH peptide. This hypothesis was tested and validated by molecular virology studies. Broad-spectrum antiviral activity of the AH peptide has been identified against HCV, HIV, herpes simplex virus, and dengue virus, and many more deadly pathogens. As a result, the AH peptide is the first in class of broad-spectrum, lipid envelope-rupturing antiviral agents, and has entered the drug pipeline. In summary, engineering strategies break down complex biological systems into simplified biomimetic models that recapitulate the most important parameters. This approach is particularly advantageous for membrane-associated biological processes because model membrane platforms provide more direct characterization of target interactions than is possible with other methods. Consequently, model membrane platforms hold great promise for solving important biomedical problems and speeding up the translation of biological knowledge into clinical applications.

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Section: Engineering Strategy To Mimic Biological Membranesmentioning

confidence: 99%

Model Membrane Platforms for Biomedicine: Case Study on Antiviral Drug Development

Jackman

Cho

2012

Biointerphases

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“…They include work by Guidelli et al on phospholipid monolayers and hybrid bilayer lipid membranes (hBLMs) on hanging-drop mercury electrodes, [32,37] by Nelson et al on phospholipid monolayers on hanging-drop mercury electrodes, [38] by Jadhav et al on hBLMs on gold electrodes [27] and the "ion channel switch biosensor" of Cornell et al, which included gramicidin covalently bound to antibody fragments in a tBLM system. [39] The findings presented here best resemble those reported by Becucci and Guidelli, [40] where the addition of gramicidin and application of negative potentials allowed the substructure of the DPTL SAM to be identified by the distribution of K + ions, as indicated by capacitance.…”

Section: à2mentioning

confidence: 99%

Effect of the Structure of Cholesterol‐Based Tethered Bilayer Lipid Membranes on Ionophore Activity

et al. 2010

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Tethered bilayer lipid membranes (tBLM) are formed on 1) pure tether lipid triethyleneoxythiol cholesterol (EO(3)C) or on 2) mixed self-assembled monolayers (SAMs) of EO(3)C and 6-mercaptohexanol (6MH). While EO(3)C is required to form a tBLM with high resistivity, 6MH dilutes the cholesterol content in the lower leaflet of the bilayer forming ionic reservoirs required for submembrane hydration. Here we show that these ionic reservoirs are required for ion transport through gramicidin or valinomycin, most likely due to the thermodynamic requirements of ions to be solvated once transported through the membrane. Unexpectedly, electrochemical impedance spectroscopy (EIS) shows an increase of capacitance upon addition of gramicidin, while addition of valinomycin decreases the membrane resistance in the presence of K(+) ions. We hypothesise that this is due to previously reported phase separation of EO(3)C and 6MH on the surface. This results in ionic reservoirs on the nanometre scale, which are not fully accounted for by the equivalent circuits used to describe the system.

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“…A performance summary of the ISA chip is presented in table 1. A tethered bilayer lipid membrane (tBLM) is useful in functional proteomics research to characterize novel membrane proteins and can be used to develop membrane protein biosensors [4,33]. The equivalent circuit model of a tBLM interface is presented in figure 10, where R m and C m represent the resistance and capacitance, respectively, of the lipid bilayer and any embedded proteins, R S is the solution resistance and C dl is the double layer capacitance [33].…”

Section: Test Resultsmentioning

confidence: 99%

High-throughput impedance spectroscopy biosensor array chip

Liu

Mason

2014

Phil. Trans. R. Soc. A.

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Impedance spectroscopy is a powerful tool for characterizing materials that exhibit a frequency dependent behaviour to an applied electric field. This paper introduces a fully integrated multi-channel impedance extraction circuit that can both generate AC stimulus signals over a broad frequency range and also measure and digitize the real and imaginary components of the impedance response. The circuit was fabricated in a 0.5 μm complementary metaloxide semiconductor. Tailored for cellular membrane interface characterization, the signal generator produces sinusoidal waves from 10 mHz to 10 kHz. To suit a variety of applications, the impedance extraction circuit provides a programmable current measurement range from 100 pA to 100 nA with a measured resolution of approximately 100 fA. Occupying only 0.045 mm 2 per measurement channel, the circuit is compact enough to include nearly 200 channels in a 3 × 3 mm 2 die area.

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Fabrication of highly insulating tethered bilayer lipid membrane using yeast cell membrane fractions for measuring ion channel activity

Cited by 38 publications

References 37 publications

Model Membrane Platforms for Biomedicine: Case Study on Antiviral Drug Development

Model Membrane Platforms for Biomedicine: Case Study on Antiviral Drug Development

Effect of the Structure of Cholesterol‐Based Tethered Bilayer Lipid Membranes on Ionophore Activity

High-throughput impedance spectroscopy biosensor array chip

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