Earticle

초록

영어

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 β-N-acetylglucosaminidase (GlcNAcase), a hexosaminidase that exists in Golgi membrane and cuts off a terminal N-acetylglucosamine (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 Nglycan patterns in Drosophila S2 cells using two suppression strategies; RNA interference (RNAi) that is a mRNA-level method and specific chemical inhibitor, 2-acetamido-1,2-dideoxynojirimycin (2-ADN), that is a protein-level method. Compared to the original N-glycan sample from human erythropoietin (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 high-performance liquid chromatography (HPLC) and matrix-assisted laser desorption/ionization time of flight mass spectrometry (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

• Hyung Joon Cha [ National Research Laboratory of Molecular Biotechnology and Department of Chemical Engineering, Pohang University of Science and Technology ]
• Yeon Kyu Kim [ National Research Laboratory of Molecular Biotechnology and Department of Chemical Engineering, Pohang University of Science and Technology ]
• Kyoung Ro Kim [ National Research Laboratory of Molecular Biotechnology and Department of Chemical Engineering, Pohang University of Science and Technology ]
• So Young Jang [ Korea Basic Science Institute ]
• Young Hwan Kim [ Korea Basic Science Institute ]

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간행물

표지보기 관심저널 등록

• 간행물명

  한국생물공학회 학술대회

• 간기

  반년간

• 수록기간

  1985~2013

• 십진분류

  KDC 476 DDC 576