期刊
JOURNAL OF BIOTECHNOLOGY
卷 153, 期 3-4, 页码 145-152出版社
ELSEVIER SCIENCE BV
DOI: 10.1016/j.jbiotec.2011.03.021
关键词
Drosophila S2 cells; N-Glycosylation; beta-N-Acetylglucosaminidase; beta 1,4-Galactosyltransferase; Human erythropoietin
资金
- Korea Research Foundation [R01-2006-000-10055-0, 2010-0001439]
- Ministry of Education, Science and Technology, Korea
- National Research Foundation of Korea [R01-2006-000-10055-0] Funding Source: Korea Institute of Science & Technology Information (KISTI), National Science & Technology Information Service (NTIS)
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.
作者
我是这篇论文的作者
点击您的名字以认领此论文并将其添加到您的个人资料中。
推荐
暂无数据