Temporary Membrane Permeabilization via the Pore-Forming Toxin Lysenin
Date
2020
Authors
Shrestha, Nisha
Thomas, Christopher A.
Richtsmeier, Devon
Bogard, Andrew
Hermann, Rebecca
Walker, Malyk
Abatchev, Gamid
Brown, Raquel J.
Fologea, Daniel
Journal Title
Journal ISSN
Volume Title
Publisher
Toxins
Abstract
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 a ect 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 e ective 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.
Description
Keywords
lysenin, permeabilization, pore forming toxins, chitosan
Citation
Shrestha, N., Thomas, C. A., Richtsmeier, D., Bogard, A., Hermann, R., Walker, M. Abatchev, G., Brown, R. J., & Fologea, D. (2020). Temporary Membrane Permeabilization via the Pore-Forming Toxin Lysenin. Toxins, 12(5), 1-17. https://doi.org/10.3390/toxins12050343.