論文 - 詳細
| RRC ID | 50440 |
|---|---|
| 著者 | Morokuma D, Xu J, Hino M, Mon H, Merzaban JS, Takahashi M, Kusakabe T, Lee JM. |
| タイトル | Expression and Characterization of Human β-1, 4-Galactosyltransferase 1 (β4GalT1) Using Silkworm-Baculovirus Expression System. |
| ジャーナル | Mol Biotechnol |
| Abstract |
Baculovirus expression vector system (BEVS) is widely known as a mass-production tool to produce functional recombinant glycoproteins except that it may not be always suitable for medical practice due to the differences in the structure of N-linked glycans between insects and mammalian. Currently, various approaches have been reported to alter N-linked glycan structures of glycoproteins derived from insects into terminally sialylated complex-type N-glycans. In the light of those studies, we also proposed in vitro maturation of N-glycan with mass-produced and purified glycosyltransferases by silkworm-BEVS. β-1,4-Galactosyltransferase 1 (β4GalT1) is known as one of type II transmembrane enzymes that transfer galactose in a β-1, 4 linkage to accepter sugars, and a key enzyme for further sialylation of N-glycans. In this study, we developed a large-scale production of recombinant human β4GalT1 (rhβ4GalT1) with N- or C-terminal tags in silkworm-BEVS. We demonstrated that rhβ4GalT1 is N-glycosylated and without mucin-type glycosylation. Interestingly, we found that purified rhβ4GalT1 from silkworm serum presented higher galactosyltransferase activity than that expressed from cultured mammalian cells. We also validated the UDP-galactose transferase activity of produced rhβ4GalT1 proteins by using protein subtracts from silkworm silk gland. Taken together, rhβ4GalT1 from silkworms can become a valuable tool for producing high-quality recombinant glycoproteins with mammalian-like N-glycans. |
| 巻・号 | 59(4-5) |
| ページ | 151-158 |
| 公開日 | 2017-5-1 |
| DOI | 10.1007/s12033-017-0003-1 |
| PII | 10.1007/s12033-017-0003-1 |
| PMID | 28342150 |
| MeSH | Animals Baculoviridae / genetics* Binding Sites Bombyx / genetics Bombyx / metabolism* Cloning, Molecular / methods Enzyme Activation Enzyme Stability Galactosyltransferases / biosynthesis* Galactosyltransferases / chemistry* Galactosyltransferases / ultrastructure Humans Protein Binding Recombinant Proteins / chemistry Recombinant Proteins / genetics Recombinant Proteins / metabolism Transfection / methods* |
| IF | 2.022 |
| 引用数 | 10 |
| オルトメトリクス指標 |
オルトメトリクス指標項目
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| 最多言及媒体 | X(Twitter) |
| 各媒体での言及数の合計 | 3 |
| 過去6か月間でのオルトメトリクス指標の変動値 | 0.0 |
| リソース情報 | |
| カイコ | n17 |