title: RNAi suppression of β-N-acetylglucosaminidase (BmFDL) for complex-type N-linked glycan synthesis in cultured silkworm cells.
abstract: Glycoproteins have various biological functions including enzymatic activity, protein stability and others. Due to the presence of paucimannosidic N-linked glycans, recombinant proteins from an insect cell expression system may not be suitable for therapeutic use. Because baculovirus expression systems (BESs) are used to produce recombinant proteins, it is of interest to modify the endogenous N-glycosylation pathway in insects to mimic that of mammals. Using a soaking RNAi sensitive cell line, BmN4-SID1, has enabled us to suppress Bombyx mori FDL (BmFDL), an N-linked glycan-specific β-N-acetylglucosaminidase. Western blotting and MALDI-TOF MS demonstrated that the BmFDL depletion almost completely converted the paucimannosidic structures of the recombinant proteins produced by BES into a complex-type structure. This highly efficient, simple and low-cost method can be used for mass production of secretion proteins with complex-type N-linked glycans.
authors.affiliation: Laboratory of Silkworm Science, Kyushu University Graduate School of Bioresource and Bioenvironmental Sciences, Fukuoka, Japan.
authors.forename: Yudai
authors.initials: Y
authors.lastname: Nagata
authors.forename: Jae Man
authors.initials: JM
authors.lastname: Lee
authors.forename: Hiroaki
authors.initials: H
authors.lastname: Mon
authors.forename: Shigeo
authors.initials: S
authors.lastname: Imanishi
authors.forename: Sun Mee
authors.initials: SM
authors.lastname: Hong
authors.forename: Shoji
authors.initials: S
authors.lastname: Komatsu
authors.forename: Yuji
authors.initials: Y
authors.lastname: Oshima
authors.forename: Takahiro
authors.initials: T
authors.lastname: Kusakabe
mesh_terms: D000118:Acetylglucosaminidase / Q000037:antagonists & inhibitors*; D000818:Animals; D012831:Bombyx / Q000166:cytology*; D002460:Cell Line; D020868:Gene Silencing; D006031:Glycosylation; D011134:Polysaccharides / Q000378:metabolism*; D011994:Recombinant Proteins / Q000378:metabolism
chemical_list: D011134:Polysaccharides; D011994:Recombinant Proteins; D000118:Acetylglucosaminidase