N-linked glycan structures of mouse interferon-beta produced by Bombyx mori larvae |
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Authors: | Misaki Ryo Nagaya Hidekazu Fujiyama Kazuhito Yanagihara Itaru Honda Takeshi Seki Tatsuji |
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Affiliation: | The International Center for Biotechnology, Osaka University, 2-1 Yamada-oka, Suita, 565-0871, Osaka, Japan. |
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Abstract: | 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. |
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Keywords: | Interferon-β N-glycan Silkworm |
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