A functional polypeptide N-acetylgalactosaminyltransferase (PGANT) initiates O-glycosylation in cultured silkworm BmN4 cells

Jian Xu, Akihiro Morio, Daisuke Morokuma, Yudai Nagata, Masato Hino, Akitsu Masuda, Zhiqing Li, Hiroaki Mon, Takahiro Kusakabe, Jae Man Lee

Research output: Contribution to journalArticle

2 Citations (Scopus)

Abstract

Mucin-type O-glycosylation is initiated by UDP-GalNAc:polypeptide N-acetylgalactosaminyltransferases (ppGalNAc-Ts or PGANTs), attaching GalNAc to serine or threonine residue of a protein substrate. In the insect model from Lepidoptera, silkworm (Bombyx mori), however, O-glycosylation pathway is totally unexplored and remains largely unknown. In this study, as the first report regarding protein O-glycosylation analysis in silkworms, we verified the O-glycan profile that a common core 1 Gal (β1-3) GalNAc disaccharide branch without terminally sialylated structure is mainly formed for a baculovirus-produced human proteoglycan 4 (PRG4) protein. Intriguingly, functional screenings in cultured silkworm BmN4 cells for nine Bmpgants reveal that Bmpgant2 is the solo functional BmPGANT for PRG4, implying that Bmpgants may have unique cell/tissue or protein substrate preferences. Furthermore, a recombinant BmPGANT2 protein was successfully purified from silkworm-BEVS and exhibited a high ability to transfer GalNAc for both peptide and protein substrates. Taken together, the present results clarified the functional BmPGANT2 in cultured silkworm cells, providing crucial fundamental insights for future studies dissecting the detailed silkworm O-glycosylation pathways and productions of glycoproteins with O-glycans.

Original languageEnglish
Pages (from-to)8783-8797
Number of pages15
JournalApplied Microbiology and Biotechnology
Volume102
Issue number20
DOIs
Publication statusPublished - Oct 1 2018

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Bombyx
Glycosylation
Polysaccharides
Proteins
Lepidoptera
Disaccharides
Baculoviridae
Mucins
Proteoglycans
Threonine
polypeptide N-acetylgalactosaminyltransferase
Recombinant Proteins
Serine
Insects
Cultured Cells
Glycoproteins
Peptides

All Science Journal Classification (ASJC) codes

  • Biotechnology
  • Applied Microbiology and Biotechnology

Cite this

A functional polypeptide N-acetylgalactosaminyltransferase (PGANT) initiates O-glycosylation in cultured silkworm BmN4 cells. / Xu, Jian; Morio, Akihiro; Morokuma, Daisuke; Nagata, Yudai; Hino, Masato; Masuda, Akitsu; Li, Zhiqing; Mon, Hiroaki; Kusakabe, Takahiro; Lee, Jae Man.

In: Applied Microbiology and Biotechnology, Vol. 102, No. 20, 01.10.2018, p. 8783-8797.

Research output: Contribution to journalArticle

Xu, Jian ; Morio, Akihiro ; Morokuma, Daisuke ; Nagata, Yudai ; Hino, Masato ; Masuda, Akitsu ; Li, Zhiqing ; Mon, Hiroaki ; Kusakabe, Takahiro ; Lee, Jae Man. / A functional polypeptide N-acetylgalactosaminyltransferase (PGANT) initiates O-glycosylation in cultured silkworm BmN4 cells. In: Applied Microbiology and Biotechnology. 2018 ; Vol. 102, No. 20. pp. 8783-8797.
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AU - Xu, Jian

AU - Morio, Akihiro

AU - Morokuma, Daisuke

AU - Nagata, Yudai

AU - Hino, Masato

AU - Masuda, Akitsu

AU - Li, Zhiqing

AU - Mon, Hiroaki

AU - Kusakabe, Takahiro

AU - Lee, Jae Man

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