N-linked glycan structures of mouse interferon-beta produced by Bombyx mori larvae.
The full-length mouse interferon-beta (mIFN-beta) cDNA, including the secretion signal peptide coding region under control of the polyhedrin promoter, was introduced into Bombyx mori nucleopolyhedrovirus (BmNPV). Recombinant mIFN-beta (rmIFN-beta) was accumulated in the haemolymph of infected silkworm larvae. Western blot analysis showed isoforms of rmIFN-beta, suggesting that rmIFN-beta is glycosylated. The glycan structures of purified rmIFN-beta were determined. The N-glycans were liberated by hydrazinolysis and the resulting oligosaccharides were labeled with 2-aminopyridine. The pyridylaminated (PA) glycans were purified by gel filtration, reversed-phase HPLC, and size-fractionation HPLC. The structures of the PA-sugar chains were identified by a combination of two-dimensional PA-sugar chain mapping, MS analysis, and exoglycosidase digestions.
Misaki
Ryo
R
The International Center for Biotechnology, Osaka University, 2-1 Yamada-oka, Suita, 565-0871, Osaka, Japan.
Nagaya
Hidekazu
H
Fujiyama
Kazuhito
K
Yanagihara
Itaru
I
Honda
Takeshi
T
Seki
Tatsuji
T
D000818:Animals; D012831:Bombyx; D016899:Interferon-beta / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D007814:Larva; D051379:Mice; D008970:Molecular Weight; D011134:Polysaccharides / Q000737:chemistry* / Q000378:metabolism*; D011487:Protein Conformation; D015202:Protein Engineering / Q000379:methods; D020033:Protein Isoforms / Q000737:chemistry / Q000378:metabolism; D017434:Protein Structure, Tertiary; D011994:Recombinant Proteins; D013045:Species Specificity; D013329:Structure-Activity Relationship
D011134:Polysaccharides; D020033:Protein Isoforms; D011994:Recombinant Proteins; D016899:Interferon-beta