| 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
|
| リソース情報 |
| カイコ |
n17 |
