Glycosaminoglycans are required for translocation of amphipathic cell-penetrating peptides across membranes.
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.
Institute of Molecular and Cell Biology, University of Tartu, Tartu, Estonia.
Janely
J
Pae
Institute of Molecular and Cell Biology, University of Tartu, Tartu, Estonia.
Laura
L
Liivamägi
Institute of Molecular and Cell Biology, University of Tartu, Tartu, Estonia.
Dmitri
D
Lubenets
Institute of Technology, University of Tartu, Tartu, Estonia.
Piret
P
Arukuusk
Institute of Technology, University of Tartu, Tartu, Estonia; Department of Neurochemistry, Stockholm University, Stockholm, Sweden.
Ülo
Ü
Langel
Institute of Molecular and Cell Biology, University of Tartu, Tartu, Estonia. Electronic address: mpooga@ut.ee.
Margus
M
Pooga
D000595:Amino Acid Sequence; D000818:Animals; D016466:CHO Cells; D057846:Cell-Penetrating Peptides / Q000378:metabolism*; D006224:Cricetinae; D003412:Cricetulus; D019004:Galanin / Q000378:metabolism; D006025:Glycosaminoglycans / Q000502:physiology*; D019763:Heparin Lyase / Q000494:pharmacology; D006801:Humans; D009842:Oligopeptides / Q000378:metabolism; D010446:Peptide Fragments / Q000378:metabolism; D021381:Protein Transport; D018342:Receptors, Adrenergic, beta-1 / Q000378:metabolism; D011993:Recombinant Fusion Proteins / Q000378:metabolism; D022161:Transport Vesicles / Q000737:chemistry / Q000378:metabolism*; D014862:Wasp Venoms / Q000378:metabolism; D014909:Wheat Germ Agglutinins; D054322:tat Gene Products, Human Immunodeficiency Virus / Q000378:metabolism
D057846:Cell-Penetrating Peptides; D006025:Glycosaminoglycans; D009842:Oligopeptides; D010446:Peptide Fragments; D018342:Receptors, Adrenergic, beta-1; D011993:Recombinant Fusion Proteins; D014862:Wasp Venoms; D014909:Wheat Germ Agglutinins; C491427:nonaarginine; D054322:tat Gene Products, Human Immunodeficiency Virus; C110068:transportan; C586078:transportan-10; D019004:Galanin; D019763:Heparin Lyase
Cell-penetrating peptide; Direct translocation; Glycosaminoglycan; Nona-arginine; Tat peptide; Transportan