| 학회 |
한국화학공학회 |
| 학술대회 |
2008년 가을 (10/23 ~ 10/24, 부산 BEXCO) |
| 권호 |
14권 2호, p.2699 |
| 발표분야 |
생물화공 |
| 제목 |
Suppression of β-N-acetylglucosaminidase in N-glycosylation Pathway for Complex Glycoprotein Formation in Drosophila S2 Cells |
| 초록 |
Most of insect cells have a simple N-glycosylation process and consequently, paucimannosidic or simple core glycans are predominant. Previously, we also revealed that paucimannosidic N-glycan structures are dominant in Drosophila S2 cells. It has been proposed that GlcNAcase, a hexosaminidase that exists in Golgi membrane and cuts off a terminal GlcNAc, might be a factor for simple N-glycosylation in several insects and their derived cells. In the present work, we investigated substantial suppression effects of GlcNAcase on N-glycan patterns in Drosophila S2 cells using two suppression strategies; RNAi that is a mRNA-level method and specific chemical inhibitor, 2-ADN, that is a protein-level method. Compared to the original N-glycan sample from hEPO-secreting stably transfected S2 cells, we found that improved N-glycan structures were clearly shown to have a terminal GlcNAc and/or galactose through HPLC and MALDI-TOF MS analyses. Therefore, we proved that GlcNAcase is a possible major factor for formation of paucimannosidic core N-glycans in Drosophila S2 cells. These data suggest that complex glycoproteins might be possible in the engineered Drosophila S2 cells by suppression of GlcNAcase in N-glycosylation pathway. |
| 저자 |
김경로1, 김연규1, 강동균1, 장소영2, 김영환2, 차형준1
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| 소속 |
1포항공과대, 2한국기초과학지원(연) |
| 키워드 |
Drosophila S2 cells; N-glycosylation; β-N-acetylglucosaminidase; Suppression; Glycoprotein; Human erythropoietin
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| E-Mail |
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| 원문파일 |
초록 보기 |
