Cell-penetrating peptide induces leaky fusion of liposomes containing late endosome-specific anionic lipid.
Cationic cell-penetrating peptides (CPPs) are a promising vehicle for the delivery of macromolecular drugs. Although many studies have indicated that CPPs enter cells by endocytosis, the mechanisms by which they cross endosomal membranes remain elusive. On the basis of experiments with liposomes, we propose that CPP escape into the cytosol is based on leaky fusion (i.e., fusion associated with the permeabilization of membranes) of the bis(monoacylglycero)phosphate (BMP)-enriched membranes of late endosomes. In our experiments, prototypic CPP HIV-1 TAT peptide did not interact with liposomes mimicking the outer leaflet of the plasma membrane, but it did induce lipid mixing and membrane leakage as it translocated into liposomes mimicking the lipid composition of late endosome. Both membrane leakage and lipid mixing depended on the BMP content and were promoted at acidic pH, which is characteristic of late endosomes. Substitution of BMP with its structural isomer, phosphatidylglycerol (PG), significantly reduced both leakage of the aqueous probe from liposomes and lipid mixing between liposomes. Although affinity of binding to TAT was similar for BMP and PG, BMP exhibited a higher tendency to support the inverted hexagonal phase than PG. Finally, membrane leakage and peptide translocation were both inhibited by inhibitors of lipid mixing, further substantiating the hypothesis that cationic peptides cross BMP-enriched membranes by inducing leaky fusion between them.
Section on Membrane Biology, Laboratory of Cellular and Molecular Biophysics, Eunice Kennedy Shriver National Institute of Child Health and Human Development, National Institutes of Health, Bethesda, Maryland, USA.
Sung-Tae
ST
Yang
Elena
E
Zaitseva
Leonid V
LV
Chernomordik
Kamran
K
Melikov
D057846:Cell-Penetrating Peptides / Q000378:metabolism*; D004396:Coloring Agents / Q000378:metabolism; D003600:Cytosol / Q000378:metabolism; D011992:Endosomes / Q000378:metabolism*; D050356:Lipid Metabolism; D008055:Lipids / Q000737:chemistry*; D008081:Liposomes / Q000378:metabolism*; D008954:Models, Biological; D010446:Peptide Fragments / Q000378:metabolism; D010539:Permeability; D010710:Phosphates / Q000737:chemistry / Q000378:metabolism; D012995:Solubility; D014867:Water / Q000737:chemistry; D054322:tat Gene Products, Human Immunodeficiency Virus / Q000737:chemistry
D057846:Cell-Penetrating Peptides; D004396:Coloring Agents; D008055:Lipids; D008081:Liposomes; D010446:Peptide Fragments; D010710:Phosphates; D054322:tat Gene Products, Human Immunodeficiency Virus; D014867:Water