バキュロウイルスが作る糖鎖リニアトリマンノースは腸管上皮細胞上に発現している

N-glycosylation of a baculovirus-expressed recombinant glycoprotein in three insect cell lines.

Kulakosky PC¹, Shuler ML, Wood HA.

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Abstract

The capacity of two Trichoplusia ni (TN-368 and BTI-Tn-5b 1-4) and a Spodoptera frugiperda (IPLB-SF-21A) cell lines to glycosylate recombinant, baculovirus-encoded, secreted, placental alkaline phosphatase was compared. The alkaline phosphatase from serum-containing, cell culture medium was purified by phosphate affinity column chromatography. The N-linked oligosaccharides were released from the purified protein with PNGase F and analyzed by fluorophore-assisted carbohydrate electrophoresis. The majority of oligosaccharide structures produced by the three cell lines contained two or three mannose residues, with and without core fucosylation, but there were structures containing up to seven mannose residues. The oligosaccharides that were qualitatively or quantitatively different between the cell lines were sequenced with glycosidase digestions. The S. frugiperda cells produced more fucosylated oligosaccharides than either of the T. ni cell lines. The smallest oligosaccharide produced by S. frugiperda cells was branched trimannose. In contrast, both T. ni cell lines produced predominantly dimannose and linear trimannose structures devoid of alpha 1-3-linked mannose.

 

 

腸管上皮細胞に発現するLypd8がとくに鞭毛をもつ細菌の腸管上皮細胞への侵入を抑制する

潰瘍性大腸炎についてはその病因として粘膜バリアの異常が注目されている

 

 

https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3647355/

While native mammalian glycoproteins often have complex type N-glycans with terminal sialic acids, insect cell-derived recombinant glycoproteins usually have much simpler side chains, known as paucimannosidic N-glycans, at sites normally occupied by complex, terminally sialylated structures. In addition, insect-derived N-glycans may contain core α1,3-linked fucose residues, which are known to be allergenic (reviewed in references [20–23]). These structural differences between the major N-glycans synthesized by insect and mammalian cells are serious problems that have hindered the use of the baculovirus-insect cell expression system for the production of recombinant glycoproteins for at least some pharmaceutical applications.