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N-Glycosylation and N-Glycan Moieties of CTB Expressed in Rice Seeds

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Abstract

Cholera toxin B subunit (CTB) is widely used as a carrier molecule and mucosal adjuvant and for the expression of fusion proteins of interest. CTB-fusion proteins are also expressed in plants, but the N-glycan structures of CTB have not been clarified. To gain insights into the N-glycosylation and N-glycans of CTB expressed in plants, we expressed CTB in rice seeds with an N-terminal glutelin signal and a C-terminal KDEL sequence and analyzed its N-glycosylation and N-glycan structures. CTB was successfully expressed in rice seeds in two forms: a form with N-glycosylation at Asn32 that included both plant-specific N-glycans and small oligomannosidic N-glycans and a non-N-glycosylated form. N-Glycan analysis of CTB showed that approximately 50 % of the N-glycans had plant-specific M3FX structures and that almost none of the N-glycans was of high-mannose-type N-glycan even though the CTB expressed in rice seeds contains a C-terminal KDEL sequence. These results suggest that the CTB expressed in rice was N-glycosylated through the endoplasmic reticulum (ER) and Golgi N-glycosylation machinery without the ER retrieval.

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Authors and Affiliations

  1. International Center for Biotechnology, Osaka University, 2-1 Yamada-oka, Suita-shi, Osaka, 565-0871, Japan

    Hiroyuki Kajiura & Kazuhito Fujiyama

  2. Pulp and Paper Research Laboratory, Nippon Paper Industries Co., Ltd., 5-21-1, Oji, Kita-ku, Tokyo, 114-0002, Japan

    Masashi Wasai & Saori Kasahara

  3. Transgenic Crop Research and Development Center, National Institute of Agrobiological Sciences, Kannondai 2-1-2, Tsukuba, Ibaraki, 305-8602, Japan

    Fumio Takaiwa

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  1. Hiroyuki Kajiura
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  2. Masashi Wasai
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  3. Saori Kasahara
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  5. Kazuhito Fujiyama
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Correspondence to Kazuhito Fujiyama.

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Kajiura, H., Wasai, M., Kasahara, S. et al. N-Glycosylation and N-Glycan Moieties of CTB Expressed in Rice Seeds. Mol Biotechnol 54, 784–794 (2013). https://doi.org/10.1007/s12033-012-9626-4

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