Multiple lipid interactions of the Sendai virus fusogenic protein.
The membrane topology of the envelope of Sendai virus was investigated using various radioactive photoactivable hydrophobic reagents: 3-(trifluoromethyl)-3-(m-[125I]iodophenyl)diazirine and the two phospholipid analogues, 1-palmitoyl-2-(2-azido-4-nitro)benzoyl-sn -glycero-3- phospho[3H]choline and 1-myristoyl-2,12-amino-(4-N-3-nitro-1-azidophenyl)dodecanoyl-sn-glycero- 3-phospho[14C]choline. The hemagglutinin-neuraminidase glycoprotein and the fusogenic (F) glycoprotein were labeled by all three probes, confirming that these proteins are integral components of the viral envelope. The labeled F glycoprotein, composed of the two subunits F1 and F2, was cleaved in situ with trypsin to yield two fragments, F32 (32 kDa) and F19 (19 kDa). F2 was not labeled by any of the probes, suggesting an external location; whereas F19 was labeled by all probes and hence contains the portion of the F glycoprotein which traverses the viral envelope. Fragment F32 reacted both with 3-(trifluoromethyl)-3-(m-[125I]iodophenyl)diazirine and with 1-palmitoyl-2-(2-azido-4-nitro)benzoyl-sn-glycero-3-phospho[3H]choline, but not with 1-myristoyl-2,12-amino-(4-N-3-nitro-1-azidophenyl)dodecanoyl-sn-glycero- 3- phospho[14C]choline. This result opens the possibility that the F glycoprotein is formed by a loop-like structure having multiple interactions with viral lipids.
D001386:Azides / Q000494:pharmacology; D001389:Azirines / Q000494:pharmacology; D006023:Glycoproteins / Q000378:metabolism; D008051:Lipid Bilayers / Q000378:metabolism; D050356:Lipid Metabolism; D008956:Models, Chemical; D008970:Molecular Weight; D010222:Parainfluenza Virus 1, Human / Q000378:metabolism*; D010449:Peptide Mapping; D010713:Phosphatidylcholines / Q000494:pharmacology; D014357:Trypsin / Q000378:metabolism; D014760:Viral Fusion Proteins / Q000378:metabolism*
D001386:Azides; D001389:Azirines; D006023:Glycoproteins; D008051:Lipid Bilayers; D010713:Phosphatidylcholines; D014760:Viral Fusion Proteins; C044850:1-myristoyl-2-(12-N-(4-azido-2-nitrophenyl)aminododecanoyl)phosphatidylcholine; C037402:1-palmitoyl-2-(2-azido-4-nitro)benzoyl-glycero-3-phosphocholine; C033174:3-(trifluoromethyl)-3-(3-iodophenyl)diazirine; D014357:Trypsin