Cell-penetrating peptides (CPPs) are promising molecules as drug carriers. However, because their uptake mainly involves endocytic mechanisms, endosomal trapping of the carrier (and drug) remains a high barrier for biomedical applications. The viral fusion mimic GALA, a pH-triggered CPP, takes advantage of the decreasing pH during endosome maturation to selectively attack endosomal membranes. Below pH 6, the sequence folds into a helix and can disrupt membranes. In this study, we show that the lipid bilayer radius-of-curvature has a negligible effect on GALA-induced leakage kinetics and that GALA remains pH responsive after inserting into a lipid membrane. The peptide can be reversibly "switched" between its inactive and active states after incorporation into the hydrophobic environment of lipid membranes, even after substantially interacting with lipid chains. This ability makes GALA-based delivery a potentially safe and efficient strategy for endosomal escape.

Cationic amphiphilic peptides have been engineered to target both Gram-positive and Gram-negative bacteria while avoiding damage to other cell types. However, the exact mechanism of how these peptides target, bind, and disrupt bacterial cell membranes is not understood. One specific peptide that has been engineered to selectively capture bacteria is WLBU2 (sequence: RRWVRRVRRWVRRVVRVVRRWVRR). It has been suggested that WLBU2 activity stems from the fact that when interacting with bacterial cell membranes the peptide assumes an α-helical structure and inserts itself into the membrane. Alternatively, in the presence of mammalian cell membranes, the peptide assumes an inert β-sheet structure. To test this hypothesis, the authors applied sum frequency generation (SFG) spectroscopy and surface tensiometry to identify the structure of WLBU2 as it interacts with model lipid monolayers that mimic mammalian and bacterial cell membranes. Model mammalian cell membranes were built upon zwitterionic 1,2-dipalmitoyl-sn-glycero-3-phosphocholine lipids while bacterial cell membranes were constructed with negatively charged 1,2-dimyristoyl-sn-glycero-3-phospho-(1'-rac-glycerol) lipids. Observed changes in surface pressure at the peptide-lipid-air interface demonstrate that the peptide has a clear binding preference toward negatively charged bacteria-like lipids. The structure of both the lipids and peptides were characterized by SFG spectra collected at the monolayer interface. Changes in monolayer structure as the peptide binds were observed by tracking the intensities of SFG vibrational modes related to the acyl chains within the lipids. Peptide structures when bound to both types of lipids were determined by SFG spectra collected within the amide I vibrational band. The SFG spectra of WLBU2 interacting with the model mammalian lipid monolayer contain two peaks near 1642 and 1678 cm indicative of an inactive β-sheet structure. SFG spectra collected from the peptide bound to a bacteria-like lipid monolayer contains just a single peak near 1651 cm which corresponds to an active α-helix structure. Combined, the tensiometry and SFG results demonstrate that WLBU2 both possesses a higher binding affinity toward and is in an active α-helix structure when bound to bacterial cell membranes.

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Development of new materials for drug delivery and biosensing requires the fine-tuning of interfacial properties. We report here the influence of the poly(ethylene glycol) (PEG) grafting density in model phospholipid monolayers on the adsorption behavior of bovine serum albumin and human fibrinogen, not only with respect to the amount of adsorbed protein, but also its orientational ordering on the surface. As expected, with increasing interfacial PEG density, the amount of adsorbed protein decreases up to the point where complete protein repellency is reached. However, at intermediate concentrations, the net orientation of adsorbed fibrinogen is highest. The different proteins respond differently to PEG, not only in the amount of protein adsorbed, but also in the manner that proteins adsorb. The results show that for specific cases, tuning the interfacial PEG concentration allows to guide the protein adsorption configuration, a feature sought after in materials for both biosensing and biomedical applications.