Membrane penetration of Sendai virus glycoproteins during the early stages of fusion with liposomes as determined by hydrophobic photoaffinity labeling.
The hydrophobic photoaffinity label 3-(trifluoromethyl)-3-(m-[125I]iodophenyl)diazirine was used to label Sendai virus proteins during fusion with cardiolipin and phosphatidylserine liposomes. Preferential labeling of the viral fusion protein during the initial stages of fusion demonstrated that this protein interacts with the hydrophobic core of the target membrane as an initiating event of virus-liposome fusion. Labeling showed time, temperature, and pH dependence consistent with earlier fluorescent measurements of fusion kinetics. The present method provides conclusive evidence supporting the hypothesis that hydrophobic interaction of the fusion protein with the target bilayer is an initial event in the fusion mechanism of viral membranes.
Novick
S L
SL
Division of Chemistry and Chemical Engineering, California Institute of Technology, Pasadena 91125.
Hoekstra
D
D
D000345:Affinity Labels; D000818:Animals; D001389:Azirines; D002308:Cardiolipins / Q000378:metabolism; D002642:Chick Embryo; D006863:Hydrogen-Ion Concentration; D008081:Liposomes / Q000378:metabolism*; D008561:Membrane Fusion; D010222:Parainfluenza Virus 1, Human / Q000502:physiology*; D010718:Phosphatidylserines / Q000378:metabolism; D012588:Scintillation Counting; D013696:Temperature; D014760:Viral Fusion Proteins / Q000378:metabolism*
D000345:Affinity Labels; D001389:Azirines; D002308:Cardiolipins; D008081:Liposomes; D010718:Phosphatidylserines; D014760:Viral Fusion Proteins; C033174:3-(trifluoromethyl)-3-(3-iodophenyl)diazirine