Voltage-controlled insertion of single α-hemolysin and <i>Mycobacterium smegmatis</i> nanopores into lipid bilayer membranes

Renner, Stephan; Bessonov, Andrey; Simmel, Friedrich C.

doi:10.1063/1.3558902

Cited by 7 publications

(3 citation statements)

References 13 publications

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“…An interesting feature of parallel multichannel bilayer arrays is the direct representation they offer of both the probabilistic nature of the membrane insertion process and the variety of protein nanopores in an RHL preparation. We have not studied this systematically, as we varied experimental conditions (lipid concentration, applied voltage, 33 RHL concentration) according to the progress of each experiment. In a database of 15 experiments with 16-fold MECAs (i.e., 240 bilayers), we thus obtained 70 insertions of one solitary, well-oriented, bona fide RHL pore.…”

Section: Resultsmentioning

confidence: 99%

Nanopore-Based Single-Molecule Mass Spectrometry on a Lipid Membrane Microarray

Ankri²,

et al. 2011

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We report on parallel high-resolution electrical single-molecule analysis on a chip-based nanopore microarray. Lipid bilayers of <20 μm diameter containing single alpha-hemolysin pores were formed on arrays of subpicoliter cavities containing individual microelectrodes (microelectrode cavity array, MECA), and ion conductance-based single molecule mass spectrometry was performed on mixtures of poly(ethylene glycol) molecules of different length. We thereby demonstrate the function of the MECA device as a chip-based platform for array-format nanopore recordings with a resolution at least equal to that of established single microbilayer supports. We conclude that devices based on MECAs may enable more widespread analytical use of nanopores by providing the high throughput and ease of operation of a high-density array format while maintaining or exceeding the precision of state-of-the-art microbilayer recordings.

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Section: Resultsmentioning

confidence: 99%

Nanopore-Based Single-Molecule Mass Spectrometry on a Lipid Membrane Microarray

Ankri²,

et al. 2011

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“…S14) (12). We added synthetic DNA channels at low concentrations (≈ 200 pM) to the cis side of the setup and applied voltage pulses to facilitate incorporation into the membrane (13, 14) (fig. S11A).…”

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confidence: 99%

Synthetic Lipid Membrane Channels Formed by Designed DNA Nanostructures

Langecker

Arnaut

Martin

et al. 2012

Science

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682

655

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We created nanometer-scale transmembrane channels in lipid bilayers using self-assembled DNA-based nanostructures. Scaffolded DNA origami was used to create a stem that penetrates and spans a lipid membrane, and a barrel-shaped cap that adheres to the membrane in part via 26 cholesterol moieties. In single-channel electrophysiological measurements, we find similarities to the response of natural ion channels, such as conductances on the order of 1 nS and channel gating. More pronounced gating was seen for mutations in which a single DNA strand of the stem protruded into the channel. In single-molecule translocation experiments, we highlight one of many potential applications of the synthetic channels, namely as single DNA molecule sensing devices.

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“…Numerous new features can be obtained by including a pore insertion probability smaller than one. In this case, the probability becomes susceptible to the electrostatic potential differences across the membrane [18]. Degradation and insertion delay can lead to oscillatory behaviour.…”

Section: Introductionmentioning

confidence: 99%

Dynamics of pore synthesis and degradation in protocells

Kubitschke

Fütterer

2012

New J. Phys.

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Liposomes have found countless applications as microreactors or for studying the evolution of protocells. However, to keep reactions ongoing, exchange with the environment is required. Based on experiments with nanopores expressed by an enclosed gene expression system, we developed a model describing the observed growth dynamics quantitatively. The model depends on one parameter only and allowed estimations of hitherto unknown parameters: the diffusion coefficient of amino acids through a single pore and the initial amino acid concentration. The long-term consequences of different degradation mechanisms are also discussed: we found a surprisingly sharp threshold deciding on the question of survival of the protocell. Contents

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Voltage-controlled insertion of single α-hemolysin and Mycobacterium smegmatis nanopores into lipid bilayer membranes

Cited by 7 publications

References 13 publications

Nanopore-Based Single-Molecule Mass Spectrometry on a Lipid Membrane Microarray

Nanopore-Based Single-Molecule Mass Spectrometry on a Lipid Membrane Microarray

Synthetic Lipid Membrane Channels Formed by Designed DNA Nanostructures

Dynamics of pore synthesis and degradation in protocells

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