Negative Dipole Potentials and Carboxylic Polar Head Groups Foster the Insertion of Cell-Penetrating Peptides into Lipid Monolayers

Cell-penetrating peptides (CPPs) are polycationic sequences of amino acids recognized as some of the most effective vehicles for delivering membrane-impermeable cargos into cells. CPPs can traverse cell membranes by direct translocation, and assessing the role of lipids on the membrane permeation process is important to convene a complete model of the CPP translocation. In this work, we focus on the biophysical basis of peptide-fatty acid interactions, analyzing how the acid-base and electrostatic properties of the lipids determine the CPP adsorption and incorporation into a Langmuir monolayer, focusing thus on the first two stages of the direct translocation mechanism. We sense the binding and insertion of the peptide into the lipid structure by measuring the changes in the surface pressure, the surface potential, and the reflectivity of the interface. We show that, beyond the presence of anionic moieties, negative dipole potentials and carboxylic polar head groups significantly promote the insertion of the peptide into the monolayer. On the basis of our results, we propose the appearance of stable CPP-lipid complexes whose kinetics of formation depends on the length of the lipids' hydrocarbon chains.