Glycosaminoglycans are required for translocation of amphipathic cell-penetrating peptides across membranes

Abstract

Cell-penetrating peptides (CPPs) are considered as one of the most promising tools to mediate the cellular delivery of various biologically active compounds that are otherwise cell impermeable. CPPs can internalize into cells via two different pathways - endocytosis and direct translocation across the plasma membrane. In both cases, the initial step of internalization requires interactions between CPPs and different plasma membrane components. Despite the extensive research, it is not yet fully understood, which of these cell surface molecules mediate the direct translocation of CPPs across the plasma- and endosomal membrane. In the present study we used giant plasma membrane vesicles (GPMVs) as a model membrane system to elucidate the specific molecular mechanisms behind the internalization and the role of cell surface glycosaminoglycans (GAGs) in the translocation of four well-known CPPs, classified as cationic (nona-arginine, Tat peptide) and amphipathic (transportan and TP10). We demonstrate here that GAGs facilitate the translocation of amphipathic CPPs, but not the internalization of cationic CPPs; and that the uptake is not mediated by a specific GAG class, but rather the overall amount of these polysaccharides is crucial for the internalization of amphipathic peptides.

Chiral Cyclobutane-Containing Cell-Penetrating Peptides as Selective Vectors for Anti-Leishmania Drug Delivery Systems

“…The hydrophobicity of amphipathic CPP [ 62 , 63 ] is crucial for their high affinity binding to lipid membranes, including those from internalized vesicles [ 64 , 65 ]. In addition, the creation of high-density positive areas in amphipatic cationic peptides favors their specific binding into anionic biopolymers, such as the cell membranes of pathogens [ 66 ].…”

Creating Supported Plasma Membrane Bilayers Using Acoustic Pressure

“…Consequently, giant plasma membrane vesicles (GPMVs) obtained from the plasma membrane of living cells have been used as an intermediate model membrane system between purely artificial membranes and the living cell [6,7]. This system has proven its usefulness in multiple fields ranging from membrane biophysics [8,9] to developmental biology [10] and drug delivery [11]. An important property of GPMVs is their capacity to exhibit lipid phase separation, allowing this phenomena to be studied in a more biologically relevant context [12][13][14][15][16][17][18].…”

Creating supported plasma membrane bilayers using acoustic pressure

“…Consequently, giant plasma membrane vesicles (GPMVs) obtained from the plasma membrane of living cells have been used as an intermediate model membrane system between purely artificial membranes and the living cell [6,7]. This system has proven its usefulness in multiple fields ranging from membrane biophysics [8,9] to developmental biology [10] and drug delivery [11]. An important property of GPMVs is their capacity to exhibit lipid phase separation, allowing this phenomena to be studied in a more biologically relevant context [12][13][14][15][16][17][18].…”