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Journal of Virology, December 1998, p. 10189-10196, Vol. 72, No. 12
Danish Veterinary Laboratory,
Received 5 May 1998/Accepted 3 September 1998
Viral hemorrhagic septicemia virus (VHSV) infections cause high
losses in cultured rainbow trout in Europe. Attempts to produce a
recombinant vaccine based on the transmembrane glycoprotein (G protein)
have indicated that proper folding is important for the antigenicity
and immunogenicity of the protein. The present study was initiated to
identify the disulfide bonds and other structural aspects relevant to
vaccine design. The N-terminal amino acid residue was identified as
being a pyroglutamic acid, corresponding to Gln21 of the primary
transcript. Peptides from endoproteinase-degraded G protein were
analyzed by mass spectrometry before and after chemical reduction, and
six disulfide bonds were identified: Cys29-Cys339, Cys44-Cys295,
Cys90-Cys132, Cys172-Cys177, Cys195-Cys265, and Cys231-Cys236. Mass
spectrometric analysis in combination with glycosidases allowed
characterization of the glycan structure of the G protein. Three of
four predicted N-linked oligosaccharides were found to be predominantly
biantennary complex-type structures. Furthermore, an O-linked glycan
near the N terminus was identified. Alignment of the VHSV G protein
with five other rhabdovirus G proteins indicates that eight cysteine
residues are situated at conserved positions. This finding suggests
that there might be some common disulfide bonding pattern among the six rhabdoviruses.
0022-538X/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.
Characterization of Intramolecular Disulfide Bonds and Secondary
Modifications of the Glycoprotein from Viral Hemorrhagic Septicemia
Virus, a Fish Rhabdovirus
*
Corresponding author. Mailing address: Danish
Veterinary Laboratory, Hangøvej 2, DK8200 Aarhus N, Denmark. Phone: 45 89 37 24 74. Fax: 45 89 37 24 70. E-mail: kej{at}svs.dk.
Journal of Virology, December 1998, p. 10189-10196, Vol. 72, No. 12
0022-538X/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.
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