The penetrating properties of the tumor homing peptide LyP-1 in model lipid membranes.

Cell-penetrating peptides (CPPs) have the property to cross the plasma membrane and enhance its permeability. CPPs were successfully used to deliver numerous cargoes such as drugs, proteins, nucleic acids, imaging and radiotherapeutic agents, gold and magnetic nanoparticles, or liposomes inside cells. Although CPPs were intensively investigated over the past 20 years, the exact molecular mechanisms of translocation across membranes are still controversial and vary from passive to active mechanisms. LyP-1 is a cyclic 9-amino-acids homing peptide that specifically binds to p32 receptors overexpressed in tumor cells. tLyP-1 peptide is the linear truncated form of LyP-1 and recognizes neuropilin (NRP) receptors expressed in glioma tumor tissue. Here, we investigate the interaction of the cyclic LyP-1 peptide and linear truncated tLyP-1 peptide with model plasma membrane in order to understand their passive, energy-independent mechanism of uptake. The experiments reveal that internalization of tLyP-1 peptides depends on membrane lipid composition. Inclusion of negatively charged phosphatidylserine (PS) or cone-shaped phosphatidylethanolamine (PE) lipids in the composition of giant unilamellar vesicles facilitates the membrane adsorption and direct penetration but without inducing pore formation in membranes. In contrast, cyclic LyP-1 peptide mostly accumulates on the membrane, with very low internalization, regardless of the lipid composition. Thus, the linear tLyP-1 peptide has enhanced penetrating properties compared with the cyclic LyP-1 peptide. Development of a mutant peptide containing higher number of arginine amino acids and preserving the homing properties of tLyP-1 may be a solution for new permeable peptides that facilitate the internalization in cells and further the endosomal escape as well.