pH-dependence of intermediate steps of membrane fusion induced by the influenza fusion peptide.
Membrane fusion mediated by the influenza-virus fusion protein is activated by low pH via a cascade of reactions. Some processes among them are irreversible, such as helix hairpin formation of the ectodomain, whereas others are reversible, such as exposure of the fusion peptide. Using this property, we attempted to dissect, in temporal order, different stages of the fusion reaction involving the fusion peptide by an acidic-neutral-acidic pH cycle. The fluorescence-quenching data indicated that both insertion depth and self-assembly are pH-reversible. In addition, lipid mixing assay was demonstrated to be arrested by neutral pH. By contrast, membrane leakage was shown to be irreversible with respect to pH. Our results, along with those from other studies on the pH-dependence of membrane fusion, are used to build a model for the virus-mediated fusion event from the perspective of pH-reversibility.
Chang
Ding-Kwo
DK
Institute of Chemistry, Academia Sinica, Taipei, Taiwan 115, Republic of China. dkc@chem.sinica.edu.tw
Cheng
Shu-Fang
SF
D009327:4-Chloro-7-nitrobenzofurazan / Q000737:chemistry; D004134:Dimyristoylphosphatidylcholine; D019267:Hemagglutinin Glycoproteins, Influenza Virus / Q000502:physiology*; D006863:Hydrogen-Ion Concentration; D009980:Influenza A virus; D008051:Lipid Bilayers / Q000737:chemistry; D008561:Membrane Fusion; D010446:Peptide Fragments / Q000502:physiology*; D010715:Phosphatidylglycerols; D012235:Rhodamines / Q000737:chemistry; D013050:Spectrometry, Fluorescence; D014760:Viral Fusion Proteins / Q000502:physiology*
D019267:Hemagglutinin Glycoproteins, Influenza Virus; D008051:Lipid Bilayers; D010446:Peptide Fragments; D010715:Phosphatidylglycerols; D012235:Rhodamines; D014760:Viral Fusion Proteins; C411888:influenza virus hemagglutinin peptide 2 (1-25); C002773:dimyristoylphosphatidylglycerol; D009327:4-Chloro-7-nitrobenzofurazan; D004134:Dimyristoylphosphatidylcholine