The fusion of artificial lipid membranes induced by the synthetic arenavirus 'fusion peptide'.
The fusing activity of the synthetic 23 amino-acid fragment (fusion peptide, FP) of the fusion protein of the Lassa arenavirus membrane was tested in a model liposomal system. The resonance energy transfer between two fluorescent phospholipid probes was monitored in order to detect dioleoylphosphatidylcholine liposome fusion induced by the peptide. Fusion rates were compared at different pH values, ionic strength and calcium concentrations. FP demonstrated fusing activity at pH 4.5-5.5, indicating that the protonated form of the FP is the active one. A transmembrane proton-gradient induced by acidification was not relevant to the fusion process, since its elimination with nigericin did not affect the FP-mediated fusion. Both Ca2+ (8 mM) and the increase of the ionic strength (1 M NaCl) inhibited liposome fusion. The efficacy of liposome fusion depended also on the lipid-to-lipid ratio. Non-linear dependence was observed at a saturation ratio of 10 mol lipid per mol peptide. A model of 'side insertion' is suggested, describing FP interaction with the membrane.
Glushakova
S E
SE
Belorussian Scientific Research Institute of Epidemiology and Microbiology, Minsk, Belarus.
Omelyanenko
V G
VG
Lukashevitch
I S
IS
Bogdanov
A A
AA
Moshnikova
A B
AB
Kozytch
A T
AT
Torchilin
V P
VP
D000595:Amino Acid Sequence; D002118:Calcium / Q000378:metabolism; D002413:Cations, Divalent; D005456:Fluorescent Dyes; D006863:Hydrogen-Ion Concentration; D007836:Lassa virus / Q000502:physiology*; D008081:Liposomes; D008969:Molecular Sequence Data; D009994:Osmolar Concentration; D010446:Peptide Fragments / Q000138:chemical synthesis / Q000502:physiology; D010714:Phosphatidylethanolamines; D014760:Viral Fusion Proteins / Q000502:physiology*
D002413:Cations, Divalent; D005456:Fluorescent Dyes; D008081:Liposomes; D010446:Peptide Fragments; D010714:Phosphatidylethanolamines; D014760:Viral Fusion Proteins; C042077:N-(7-nitrobenz-2-oxa-1,3-diazol-4-yl)phosphatidylethanolamine; D002118:Calcium