NR ATTW
AU Magzoub,M.; Pramanik,A.; Gräslund,A.
TI Modeling the endosomal escape of cell-penetrating peptides: transmembrane pH gradient driven translocation across phospholipid bilayers.
QU Biochemistry 2005 Nov 15; 44(45): 14890-7
PT journal article
AB Cell-penetrating peptides (CPPs) are able to mediate the efficient cellular uptake of a wide range of cargoes. Internalization of a number of CPPs requires uptake by endocytosis, initiated by binding to anionic cell surface heparan sulfate (HS), followed by escape from endosomes. To elucidate the endosomal escape mechanism, we have modeled the process for two CPPs: penetratin (pAntp) and the N-terminal signal peptide of the unprocessed bovine prion protein (bPrPp). Large unilamellar phospholipid vesicles (LUVs) were produced encapsulating either peptide, and an ionophore, nigericin, was used to create a transmembrane pH gradient (DeltapH(mem), inside acidic) similar to the one arising in endosomes in vivo. In the absence of DeltapH(mem), no pAntp escape from the LUVs is observed, while a fraction of bPrPp escapes. In the presence of DeltapH(mem), a significant amount of pAntp escapes and an even higher degree of bPrPp escape takes place. These results, together with the differences in kinetics of escape, indicate different escape mechanisms for the two peptides. A minimum threshold peptide concentration exists for the escape of both peptides. Coupling of the peptides to a cargo reduces the fraction escaping, while complexation with HS significantly hinders the escape. Fluorescence correlation spectroscopy results show that during the escape process the LUVs are intact. Taken together, these results suggest a model for endosomal escape of CPPs: DeltapH(mem)-mediated mechanism, following dissociation from HS of the peptides, above a minimum threshold peptide concentration, in a process that does not involve lysis of the vesicles.
MH Amino Acid Sequence; Animals; Biological Transport; Carrier Proteins/chemistry/metabolism; Cattle; Cell Membrane Permeability; *Endocytosis; Endosomes/*metabolism; Heparitin Sulfate/chemistry; Hydrogen-Ion Concentration; Ionophores/pharmacology; Lipid Bilayers/metabolism; *Models, Biological; Molecular Sequence Data; Nigericin/pharmacology; Peptides/*metabolism; Phospholipids/*metabolism; Prions/chemistry/metabolism; Protons; Research Support, Non-U.S. Gov't
AD Department of Biochemistry and Biophysics, Stockholm University, S-106 91 Stockholm, Sweden.
SP englisch
PO USA