N-acetyl-beta-glucosaminidase accounts for differences in glycosylation of influenza virus hemagglutinin expressed in insect cells from a baculovirus vector
N-acetyl-beta-glucosaminidase accounts for differences in glycosylation of influenza virus hemagglutinin expressed in insect cells from a baculovirus vector
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N-乙酰-β-氨基葡萄糖苷酶解释了杆状病毒载体昆虫细胞中表达的流感病毒血凝素的糖基化差异
作者：
R. Wagner;H. Geyer;R. Geyer;H. Klenk
鸡瘟病毒的血凝素已通过使用杆状病毒载体在草地贪夜蛾（Sf9）细胞和Estigmene acrea细胞中表达。结构分析表明，来自 Sf9 细胞的 HA 的内切 H 抗性 N-聚糖主要是三甘露糖基核心寡糖，而在 E. acrea 细胞中，大多数这些核心被至少一个末端 N-乙酰葡糖胺残基延长。为了了解碳水化合物结构的差异，对参与 N-聚糖加工的酶进行了分析。结果表明，观察到的不同糖基化模式是由于 Sf9 细胞中发现的 N-乙酰基-β-氨基葡萄糖苷酶活性所致，而 E. acrea 细胞中则没有。该酶专门使用 GlcNAcMan(3)GlcNAc(2) 寡糖作为底物。当N-乙酰-β-氨基葡萄糖苷酶或α-甘露糖苷酶II受到特定抑制剂抑制时，Sf9细胞血凝素中末端N-乙酰氨基葡萄糖的量显着增加。这些结果表明，两种细胞系中的 N 糖基化均遵循经典途径直至 GlcNAcMan(3)GlcNAc(2) 寡糖侧链阶段。虽然这些结构是 E. acrea 细胞中的最终产物，但它们在 Sf9 细胞中被 N-乙酰基-β-氨基葡萄糖苷酶降解为 Man(3)GlcNAc(2) 核心。
The hemagglutinin of fowl plague virus has been expressed in Spodoptera frugiperda (Sf9) cells and in Estigmene acrea cells by using a baculovirus vector. Structural analysis revealed that the endo-H-resistant N-glycans of HA from Sf9 cells were predominantly trimannosyl core oligosaccharides, whereas in E. acrea cells most of these cores were elongated by at least one terminal N-acetylglucosamine residue. To understand the difference in carbohydrate structures, enzymes involved in N-glycan processing have been analyzed. The results revealed that the different glycosylation patterns observed are due to an N-acetyl-beta-glucosaminidase activity that was found in Sf9 cells but not in E. acrea cells. This enzyme specifically used the GlcNAcMan(3)GlcNAc(2) oligosaccharide as a substrate. When N-acetyl-beta-glucosaminidase or alpha-mannosidase II was inhibited by specific inhibitors, the amount of terminal N-acetylglucosamine in hemagglutinin from Sf9 cells was significantly enhanced. These results demonstrate that N glycosylation in both cell lines follows the classical pathway up to the stage of GlcNAcMan(3)GlcNAc(2) oligosaccharide side chains. Whereas these structures are the end product in E. acrea cells, they are degraded in Sf9 cells to Man(3)GlcNAc(2) cores by N-acetyl-beta-glucosaminidase.
天冬酰胺连接的寡糖加工的调节。
DOI：
--
发表时间：
1984
期刊：
The Journal of biological chemistry
影响因子：
--
作者：
Hsieh,P;Robbins,PW
通讯作者：
Robbins,PW
鳞翅目昆虫（草地贪夜蛾）细胞表达人纤溶酶原 cDNA 时的低聚糖加工。
影响因子：
2.9
作者：
Davidson,DJ;Fraser,MJ;Castellino,FJ
通讯作者：
Castellino,FJ
草地贪夜蛾细胞 (IPLB-SF-21AE) 中存在 UDP-N-乙酰氨基葡萄糖:α-3-D-甘露糖苷 β 1,2-N-乙酰氨基葡萄糖转移酶 I 活性及其因杆状病毒感染而增强。
DOI：
--
发表时间：
1993
期刊：
The Journal of biological chemistry
影响因子：
--
作者：
Velardo,MA;Bretthauer,RK;Boutaud,A;Reinhold,B;Reinhold,VN;Castellino,FJ
通讯作者：
Castellino,FJ