Fusion of Sendai virus with vesicles of oligomerizable lipids: a microcalorimetric analysis of membrane fusion.
Sendai virus fuses efficiently with small and large unilamellar vesicles of the lipid 1,2-di-n-hexadecyloxypropyl-4- (beta-nitrostyryl) phosphate (DHPBNS) at pH 7.4 and 37 degrees C, as shown by lipid mixing assays and electron microscopy. However, fusion is strongly inhibited by oligomerization of the head groups of DHPBNS in the bilayer vesicles. The enthalpy associated with fusion of Sendai virus with DHPBNS vesicles was measured by isothermal titration microcalorimetry, comparing titrations of Sendai virus into (i) solutions of DHPBNS vesicles (which fuse with the virus) and (ii) oligomerized DHPBNS vesicles (which do not fuse with the virus), respectively. The observed heat effect of fusion of Sendai virus with DHPBNS vesicles is strongly dependent on the buffer medium, reflecting a partial charge neutralization of the Sendai F and HN proteins upon insertion into the negatively-charged vesicle membrane. No buffer effect was observed for the titration of Sendai virus into oligomerized DHPBNS vesicles, indicating that inhibition of fusion is a result of inhibition of insertion of the fusion protein into the target membrane. Fusion of Sendai virus with DHPBNS vesicles is endothermic and entropy-driven. The positive enthalpy term is dominated by heat effects resulting from merging of the protein-rich viral envelope with the lipid vesicle bilayers rather than by the fusion of the viral with the vesicle bilayers per se.
Ravoo
B J
BJ
Department of Organic and Molecular Inorganic Chemistry, University of Groningen, Nijenborgh 4, Groningen, 9747 AG, The Netherlands.
Weringa
W D
WD
Engberts
J B
JB
D002151:Calorimetry / Q000379:methods; D002462:Cell Membrane / Q000821:virology; D008055:Lipids / Q000737:chemistry / Q000276:immunology*; D008561:Membrane Fusion / Q000502:physiology*; D008563:Membrane Lipids / Q000378:metabolism*; D008854:Microscopy, Electron / Q000379:methods; D015394:Molecular Structure; D018108:Respirovirus / Q000378:metabolism* / Q000502:physiology / Q000648:ultrastructure; D017186:Titrimetry; D014760:Viral Fusion Proteins / Q000378:metabolism*
C421012:1,2-di-n-hexadecyloxypropyl-4-(beta-nitrostyryl)phosphate; D008055:Lipids; D008563:Membrane Lipids; D014760:Viral Fusion Proteins