Enhanced efficiency of a targeted fusogenic peptide.
Membrane targeting was investigated as a potential strategy to increase the fusogenic activity of an isolated fusion peptide. This was achieved by coupling the fusogenic carboxy-terminal part of the beta-amyloid peptide (Abeta, amino acids 29-40), involved in Alzheimer's disease, to a positively charged peptide (PIP2-binding peptide, PBP) interacting specifically with a naturally occurring negatively charged phospholipid, phosphatidylinositol 4, 5-bisphosphate (PIP2). Peptide-induced vesicle fusion was spectroscopically evidenced by: (i) mixing of membrane lipids, (ii) mixing of aqueous vesicular contents, and (iii) an irreversible increase in vesicle size, at concentrations five to six times lower than the Abeta(29-40) peptide. In contrast, at these concentrations the PBP-Abeta(29-40) peptide did not display any significant activity on neutral vesicles, indicating that negatively charged phospholipids included as targets in the membranes, are required to compensate for the lower hydrophobicity of this peptide. When the alpha-helical structure of the chimeric peptide was induced by dissolving it in trifluoroethanol, an increase of the fusogenic potential of the peptide was observed, supporting the hypothesis that the alpha-helical conformation of the peptide is crucial to trigger the lipid-peptide interaction. The specificity of the interaction between PIP2 and the PBP moiety, was shown by the less efficient targeting of the chimeric peptide to membranes charged with phosphatidylserine. These data thus demonstrate that the specific properties of both the Abeta(29-40) and the PBP peptide are conserved in the chimeric peptide, and that a synergetic effect is reached through chemical linkage of these two fragments.
Decout
A
A
Laboratory for Lipoprotein Chemistry, Department of Biochemistry, Universiteit Gent, B-9000 Gent, Belgium.
Labeur
C
C
Goethals
M
M
Brasseur
R
R
Vandekerckhove
J
J
Rosseneu
M
M
D004305:Dose-Response Relationship, Drug; D008081:Liposomes / Q000737:chemistry / Q000378:metabolism; D008561:Membrane Fusion / Q000187:drug effects*; D010446:Peptide Fragments / Q000235:genetics / Q000378:metabolism / Q000494:pharmacology*; D010713:Phosphatidylcholines / Q000737:chemistry / Q000378:metabolism; D010714:Phosphatidylethanolamines / Q000737:chemistry / Q000378:metabolism; D011485:Protein Binding; D011487:Protein Conformation; D017433:Protein Structure, Secondary; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000378:metabolism / Q000494:pharmacology
D008081:Liposomes; D010446:Peptide Fragments; D010713:Phosphatidylcholines; D010714:Phosphatidylethanolamines; D011993:Recombinant Fusion Proteins