Authors:

Emmaleigh Shinno

Date Created:

Not specified

Course Title:

Professor:

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About Paper:

Cationic amphiphilic polyproline helices (CAPHs) are cell penetrating peptides (CPPs) with great promise as a vehicle for drug delivery or direct bacterial killing within a cell. To maximize CAPH utility, a molecular understanding of binding and activity thresholds is needed. Structure-function relationships have been established for some CPPs but the kinetics of individual peptides interacting with the membrane may provide new insights to improve targeting and dosing. Single molecule binding on and off rates can be measured using in vitro reconstitution using glass-supported lipid bilayers (SLBs) of controlled composition. Fluorescently tagged CAPHs interacting with the membrane can be visualized with high spatial and temporal resolution in a total internal reflection fluorescence (TIRF) configuration that is membrane selective. Lipid charge, character, and densities can all be modulated and stacked supported SLBs can be used to measure the extent of membrane penetration. Some CAPHs are more cationic than others and this investigation focuses on a +9 peptide that is strongly membrane penetrating. We show that this CAPH is sensitive to membrane cholesterol suggesting that this membrane component impacts efficacy of CAPH delivery. Ongoing efforts are focused on modulating CAPH charge and increasing complexity of model membranes to recapitulate more realistic cell surfaces.

Source:

Purdue University / 2023

Topics:

No topics listed

Co-authors:

Emmaleigh Shinno