Membrane fusion activity of tick-borne encephalitis virus and recombinant subviral particles in a liposomal model system.
We present a kinetic analysis of the membrane fusion activity of tick-borne encephalitis (TBE) virus and TBE-derived recombinant subviral particles (RSPs) in a liposomal model system. Fusion was monitored using a fluorescence assay involving pyrene-labeled phospholipids. Fusion was strictly dependent on low pH, with the optimum being at pH 5.3-5.5 and the threshold at pH 6.8. Fusion did not require a protein or carbohydrate receptor in the target liposomes. Preexposure to low pH of the virus alone resulted in inactivation of its fusion activity. At the optimum pH for fusion and 37 degrees C, the rate and extent of fusion were very high, with more than 50% of the virus fusing within 2 s and the final extent of fusion being 70%. Lowering of the temperature did not result in a significant decrease in the rate and extent of fusion, suggesting that TBE virus fusion is a facile process with a low activation energy, possibly due to the flat orientation of the E glycoprotein on the viral surface facilitating the establishment of direct intermembrane contact. The fusion characteristics of TBE virus and RSPs were similar, indicating that RSPs provide a reliable and convenient model for further study of the membrane fusion properties of TBE virus.
Corver
J
J
Laboratory of Molecular Virology, University of Groningen and Academic Hospital, Groningen, Ant. Deusinglaan 1, 9713 AV, The Netherlands.
Ortiz
A
A
Allison
S L
SL
Schalich
J
J
Heinz
F X
FX
Wilschut
J
J
D000818:Animals; D002478:Cells, Cultured; D002642:Chick Embryo; D002784:Cholesterol / Q000378:metabolism; D004669:Encephalitis Viruses, Tick-Borne / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D005456:Fluorescent Dyes / Q000378:metabolism; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D008081:Liposomes / Q000737:chemistry / Q000378:metabolism*; D008561:Membrane Fusion; D008562:Membrane Glycoproteins / Q000235:genetics / Q000378:metabolism; D008954:Models, Biological; D010713:Phosphatidylcholines / Q000378:metabolism; D010714:Phosphatidylethanolamines / Q000378:metabolism; D011721:Pyrenes / Q000378:metabolism; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism; D013109:Sphingomyelins / Q000378:metabolism; D013816:Thermodynamics; D014759:Viral Envelope Proteins / Q000235:genetics / Q000378:metabolism; D015678:Viral Structural Proteins / Q000235:genetics / Q000378:metabolism*
D005456:Fluorescent Dyes; D008081:Liposomes; D008562:Membrane Glycoproteins; D010713:Phosphatidylcholines; D010714:Phosphatidylethanolamines; D011721:Pyrenes; D011994:Recombinant Proteins; D013109:Sphingomyelins; D014759:Viral Envelope Proteins; D015678:Viral Structural Proteins; D002784:Cholesterol; C030984:pyrene