Assignment of disulfide bridges in the fusion glycoprotein of Sendai virus.
The mature fusion (F) glycoprotein of the paramyxovirus family consists of two disulfide-linked subunits, the N-terminal F2 and the C-terminal F1 subunits, and contains 10 cysteine residues which are highly conserved at specific positions. The high level of conservation strongly suggests that they are indeed disulfide linked and play important roles in the folding and functioning of the molecule. However, it has not even been clarified which cysteine residues link the F2 and F1 subunits. This report describes our assignment of the disulfide bridges in purified Sendai virus F glycoprotein by fragmentation of the polypeptide and isolation of cystine-containing peptides and determination of their N-terminal sequences. The data demonstrate that all of the 10 cysteine residues participate in disulfide bridges and that Cys-70, the only cysteine in F2, and Cys-199, the most upstream cysteine in F1, form the interchain bond. Of the remaining eight cysteine residues clustered near the transmembrane domain of F1, the specific bridges identified are Cys-338 to Cys-347 and Cys-362 to Cys-370. Although no exact pairings between the subsequent four residues were defined, it seems likely that the most downstream, Cys-424, is linked to Cys-394, Cys-399, or Cys-401. Thus, we conclude that the cysteine-rich domain indeed contributes to the formation of a bunched structure containing at least two tandem cystine loops.
Iwata
S
S
Research Institute for Disease Mechanism and Control, Nagoya University School of Medicine, Japan.
Schmidt
A C
AC
Titani
K
K
Suzuki
M
M
Kido
H
H
Gotoh
B
B
Hamaguchi
M
M
Nagai
Y
Y
D000595:Amino Acid Sequence; D000596:Amino Acids / Q000032:analysis; D003553:Cystine; D004220:Disulfides; D008969:Molecular Sequence Data; D010222:Parainfluenza Virus 1, Human / Q000737:chemistry*; D010446:Peptide Fragments / Q000737:chemistry; D011487:Protein Conformation; D017421:Sequence Analysis; D014760:Viral Fusion Proteins / Q000737:chemistry*; D014771:Virion / Q000737:chemistry
D000596:Amino Acids; D004220:Disulfides; D010446:Peptide Fragments; D014760:Viral Fusion Proteins; D003553:Cystine