Fluorescence evidence for a loose self-assembly of the fusion peptide of influenza virus HA2 in the lipid bilayer.
Steady state fluorescence experiments were performed on a 25-mer synthetic peptide incorporated in the phospholipid vesicle to study the role of oligomerization of the fusion peptide in membrane fusion. It was found from fluorescence resonance energy transfer (FRET) that the extent of lipid mixing and the initial mixing rate varied with the fusion peptide concentration in a higher than linear fashion, indicating that the peptide promoted membrane mixing as oligomers. Results of self-quenching of the Rhodamine (Rho) in Rho-labelled peptide incorporated in the phospholipid bilayer indicated that the peptide molecules assembled in the bilayer with an order higher than dimer. The data also revealed that the peptides were not tightly packed in the membrane. Binding affinity measurement monitored by the NBD fluorescence intensity on the fluorophore-labelled fusion peptide supports the notion of self-association of the peptide in the vesicular dispersion. In the sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE) experiments, a diffuse band with apparent molecular mass close to a dimeric species of the wild type fusion peptide suggested that the fusion peptides formed loose oligomers under the influence of SDS detergent in the electric field. The result is in contrast to a less fusion-active variant which appears to exhibit less propensity for self-association.
Institute of Chemistry, Academia Sinica, Taipei, Taiwan, Republic of China 115.
Shu-Fang
SF
Cheng
Assen B
AB
Kantchev
Ding-Kwo
DK
Chang
D005453:Fluorescence; D031541:Fluorescence Resonance Energy Transfer; D019267:Hemagglutinin Glycoproteins, Influenza Virus / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D008051:Lipid Bilayers / Q000378:metabolism*; D050356:Lipid Metabolism; D012235:Rhodamines / Q000378:metabolism; D014760:Viral Fusion Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*
D019267:Hemagglutinin Glycoproteins, Influenza Virus; D008051:Lipid Bilayers; D012235:Rhodamines; D014760:Viral Fusion Proteins