Stochastic sensing of Angiotensin II with lysenin channels

Shrestha, Nisha; Bryant, Sheenah; Thomas, Christopher A.; Richtsmeier, Devon; Pu, Xinzhu; Tinker, Juliette; Fologea, Daniel

doi:10.1038/s41598-017-02438-0

Cited by 9 publications

(16 citation statements)

References 58 publications

(111 reference statements)

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“…Numerous nanopores of biological origin were investigated for sensing applications, such as α-hemolysin, aerolysin, E. coli ClyA toxin, lysenin, and motor proteins [5,6,8,[29][30][31][32][33][34][35][36]. Among those biological tools, lysenin channels are attractive candidates for sensor development owing to their commercial availability, facile reconstitution into artificial membranes, extended stability, intrinsic regulatory mechanisms, and a large unobstructed opening.…”

Section: Introductionmentioning

confidence: 99%

Lysenin Channels as Sensors for Ions and Molecules

Bogard

Abatchev

Hutchinson

et al. 2020

Sensors

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Lysenin is a pore-forming protein extracted from the earthworm Eisenia fetida, which inserts large conductance pores in artificial and natural lipid membranes containing sphingomyelin. Its cytolytic and hemolytic activity is rather indicative of a pore-forming toxin; however, lysenin channels present intricate regulatory features manifested as a reduction in conductance upon exposure to multivalent ions. Lysenin pores also present a large unobstructed channel, which enables the translocation of analytes, such as short DNA and peptide molecules, driven by electrochemical gradients. These important features of lysenin channels provide opportunities for using them as sensors for a large variety of applications. In this respect, this literature review is focused on investigations aimed at the potential use of lysenin channels as analytical tools. The described explorations include interactions with multivalent inorganic and organic cations, analyses on the reversibility of such interactions, insights into the regulation mechanisms of lysenin channels, interactions with purines, stochastic sensing of peptides and DNA molecules, and evidence of molecular translocation. Lysenin channels present themselves as versatile sensing platforms that exploit either intrinsic regulatory features or the changes in ionic currents elicited when molecules thread the conducting pathway, which may be further developed into analytical tools of high specificity and sensitivity or exploited for other scientific biotechnological applications.

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Section: Introductionmentioning

confidence: 99%

Lysenin Channels as Sensors for Ions and Molecules

Bogard

Abatchev

Hutchinson

et al. 2020

Sensors

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“…The limited size of the lysenin channel opening in comparison to other toxins (i.e., SLO [16]) might potentially restrict the proposed methodology. Nonetheless, ions, dyes, small peptides, and even single stranded nucleic acid molecules diffuse through the open lysenin channel [32,52], which is anticipated to lead to numerous scientific, biotechnological, and biomedical applications that are based on this controlled permeabilization approach.…”

Section: Discussionmentioning

confidence: 99%

Temporary Membrane Permeabilization via the Pore-Forming Toxin Lysenin

Shrestha

Thomas

Richtsmeier

et al. 2020

Toxins

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Pore-forming toxins are alluring tools for delivering biologically-active, impermeable cargoes to intracellular environments by introducing large conductance pathways into cell membranes. However, the lack of regulation often leads to the dissipation of electrical and chemical gradients, which might significantly affect the viability of cells under scrutiny. To mitigate these problems, we explored the use of lysenin channels to reversibly control the barrier function of natural and artificial lipid membrane systems by controlling the lysenin’s transport properties. We employed artificial membranes and electrophysiology measurements in order to identify the influence of labels and media on the lysenin channel’s conductance. Two cell culture models: Jurkat cells in suspension and adherent ATDC5 cells were utilized to demonstrate that lysenin channels may provide temporary cytosol access to membrane non-permeant propidium iodide and phalloidin. Permeability and cell viability were assessed by fluorescence spectroscopy and microscopy. Membrane resealing by chitosan or specific media addition proved to be an effective way of maintaining cellular viability. In addition, we loaded non-permeant dyes into liposomes via lysenin channels by controlling their conducting state with multivalent metal cations. The improved control over membrane permeability might prove fruitful for a large variety of biological or biomedical applications that require only temporary, non-destructive access to the inner environment enclosed by natural and artificial membranes.

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“…The experimental setup employed the use of a planar BLM chamber consisting of two polytetrafluoroethylene (PTFE) reservoirs separated by a thin (~ 120 μm) PTFE film that had been pierced with an electric spark to create a circular hole of ~ 70 µm diameter [ 45 , 46 ]. The reservoirs were filled with 1 mL buffered electrolyte and connected via two Ag/AgCl electrodes inserted in the solution to an Axopatch 200B amplifier (molecular devices).…”

Section: Methodsmentioning

confidence: 99%

ZnO nanoparticles modulate the ionic transport and voltage regulation of lysenin nanochannels

et al. 2017

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BackgroundThe insufficient understanding of unintended biological impacts from nanomaterials (NMs) represents a serious impediment to their use for scientific, technological, and medical applications. While previous studies have focused on understanding nanotoxicity effects mostly resulting from cellular internalization, recent work indicates that NMs may interfere with transmembrane transport mechanisms, hence enabling contributions to nanotoxicity by affecting key biological activities dependent on transmembrane transport. In this line of inquiry, we investigated the effects of charged nanoparticles (NPs) on the transport properties of lysenin, a pore-forming toxin that shares fundamental features with ion channels such as regulation and high transport rate.ResultsThe macroscopic conductance of lysenin channels greatly diminished in the presence of cationic ZnO NPs. The inhibitory effects were asymmetrical relative to the direction of the electric field and addition site, suggesting electrostatic interactions between ZnO NPs and a binding site. Similar changes in the macroscopic conductance were observed when lysenin channels were reconstituted in neutral lipid membranes, implicating protein-NP interactions as the major contributor to the reduced transport capabilities. In contrast, no inhibitory effects were observed in the presence of anionic SnO2 NPs. Additionally, we demonstrate that inhibition of ion transport is not due to the dissolution of ZnO NPs and subsequent interactions of zinc ions with lysenin channels.ConclusionWe conclude that electrostatic interactions between positively charged ZnO NPs and negative charges within the lysenin channels are responsible for the inhibitory effects on the transport of ions. These interactions point to a potential mechanism of cytotoxicity, which may not require NP internalization.Electronic supplementary materialThe online version of this article (10.1186/s12951-017-0327-9) contains supplementary material, which is available to authorized users.

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Stochastic sensing of Angiotensin II with lysenin channels

Cited by 9 publications

References 58 publications

Lysenin Channels as Sensors for Ions and Molecules

Lysenin Channels as Sensors for Ions and Molecules

Temporary Membrane Permeabilization via the Pore-Forming Toxin Lysenin

ZnO nanoparticles modulate the ionic transport and voltage regulation of lysenin nanochannels

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