Temporary Membrane Permeabilization via the Pore-Forming Toxin Lysenin

Area of Science:

• Membrane biophysics
• Cellular biology
• Biotechnology

Background:

• Pore-forming toxins create cell membrane channels for intracellular delivery.
• Uncontrolled pore formation can disrupt cell gradients and viability.
• Lysenin channels offer potential for regulated membrane permeability.

Purpose of the Study:

• To explore reversible control of membrane barrier function using lysenin channels.
• To investigate lysenin channel properties in natural and artificial lipid membranes.
• To demonstrate temporary cytosol access for non-permeant molecules and maintain cell viability.

Main Methods:

• Utilized artificial membranes and electrophysiology to study lysenin channel conductance.
• Employed Jurkat and ATDC5 cell culture models.
• Assessed permeability and viability using fluorescence spectroscopy, microscopy, and resealing techniques.

Main Results:

• Identified factors influencing lysenin channel conductance (labels, media).
• Demonstrated temporary cytosol access to Jurkat and ATDC5 cells for propidium iodide and phalloidin.
• Showcased effective membrane resealing with chitosan or specific media, maintaining cell viability.
• Successfully loaded non-permeant dyes into liposomes using lysenin channels controlled by metal cations.

Conclusions:

• Lysenin channels provide a method for reversible control of membrane permeability.
• Temporary, non-destructive access to cellular or liposomal interiors is achievable.
• This approach has broad potential for biological and biomedical applications requiring transient membrane access.