Expression of beta-1,4-galactosyltransferase and suppression of beta-N-acetylglucosaminidase to aid synthesis of complex N-glycans in insect Drosophila S2 cells
SCIE
SCOPUS
- Title
- Expression of beta-1,4-galactosyltransferase and suppression of beta-N-acetylglucosaminidase to aid synthesis of complex N-glycans in insect Drosophila S2 cells
- Authors
- Kim, YK; Kim, KR; Kang, DG; Jang, SY; Kim, YH; Cha, HJ
- Date Issued
- 2011-05-20
- Publisher
- ELSEVIER SCIENCE BV
- Abstract
- Previously, we have shown that simple paucimannosidic N-glycan structures in insect Drosophila S2 cells arise mainly because of beta-N-acetylglucosaminidase (GlcNAcase) action. Thus, in an earlier report, we suppressed GlcNAcase activity and clearly demonstrated that more complexN-glycans with two terminal N-acetylglucosamine (GlcNAc) residues were then synthesized. In the present work, we investigated the synergistic effects of beta-1,4-galactosyltransferase (GalT) expression and GlcNAcase suppression on N-glycan patterns. We found that the N-glycan pattern of human erythropoietin secreted by engineered S2 cells expressing GalT but not GlcNAcase was complete, even in small portion, except for sialylation; the N-glycan structures had two terminal galactose (Gal) residues. When GalT was expressed but GlcNAcase was not inhibited, N-glycan with GlcNAc and Gal at only one branch end was synthesized. Therefore, it will be possible to express a complete functional human glycoprotein in engineered Drosophila S2 cells by suppressing GlcNAcase and co-expressing additional glycosyltransferases of N-glycosylation pathway. (C) 2011 Elsevier B.V. All rights reserved.
- Keywords
- Drosophila S2 cells; N-Glycosylation; beta-N-Acetylglucosaminidase; beta 1,4-Galactosyltransferase; Human erythropoietin; SECRETED HUMAN ERYTHROPOIETIN; STATISTICAL OPTIMIZATION; GLYCOSYLATION PATHWAY; PROTEIN; GLYCOPROTEIN; MEMBRANE
- URI
- https://oasis.postech.ac.kr/handle/2014.oak/17380
- DOI
- 10.1016/J.JBIOTEC.2011.03.021
- ISSN
- 0168-1656
- Article Type
- Article
- Citation
- JOURNAL OF BIOTECHNOLOGY, vol. 153, no. 3, page. 145 - 152, 2011-05-20
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