Potential Use of Lysenin Channels in Drug Release from Liposomes

Student: Jess Ray

Major Professor: Dr. Gregory Salamo

Research Area(s):

Nanoscience and Engineering

Wild type lysenin channels were investigated under the influence of different multivalent ions.
Lysenin channels inserted into liposomes could create new opportunities for drug delivery systems.

Use the mutated lysenin channels in experiments to determine the voltage at which they gate and compare it to wild type lysenin.
Control the drug release out of the lysenin-liposome drug carriers to enhance drug delivery applications.

Use Teflon chambers to create a bilayer lipid membrane (BLM) model.
Create lipid solution containing cholesterol, sphingomyelin, and asolectin to create a model cell like animal cell membranes.
Insert lysenin in the BLM to experimentally determine gating voltages (using Axo-Patch).
Introduce multivalent ions to observe the ligand – induced gating.

Wild type lysenin channels gates at roughly 20mV, while the mutated gated at roughly 30-35mV (as the figure on the left shows).
Iron divalent cations inhibit the macroscopic current of both wild and mutated lysenin channels at low monovalent ions concentration (as the figure on the right shows).

Mutated type lysenin gated at a higher voltage than the wild type lysenin did, due to the mutation. This lead to roughly a 10-15 mV higher voltage required to cause gating in the mutated type.
Using a less concentrated monovalent solution (50mM KCl instead of 150mM) allowed the conductance of both types of lysenin to reach zero.
The charge of the mutated lysenin was reduced by the mutation. This lead to a higher concentration of iron divalent ions being needed for ligand-induced gating, as hypothesized.