BIOCHEMICAL-ANALYSIS OF THE N-GLYCOSYLATION PATHWAY IN BACULOVIRUS-INFECTED LEPIDOPTERAN INSECT CELLS
BIOCHEMICAL-ANALYSIS OF THE N-GLYCOSYLATION PATHWAY IN BACULOVIRUS-INFECTED LEPIDOPTERAN INSECT CELLS
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DOI：
10.1006/viro.1995.1508
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发表时间：
1995-10-01
期刊：
影响因子：
3.7
通讯作者：
FINN, EE
中科院分区：
文献类型：
--
作者：
JARVIS, DL;FINN, EE
杆状病毒-昆虫细胞系统通常用于外源糖蛋白的生产，但该系统中N-糖基化途径的确切性质仍不清楚。一些研究表明，这些细胞不能处理N-连接的寡糖含有外链半乳糖和唾液酸的复杂形式，而其他人表明，他们可以，在这项研究中，我们使用的杆状病毒苜蓿银纹夜蛾多衣壳核型多角体病毒（AcMNPV）的主要病毒粒子包膜糖蛋白探测杆状病毒感染的鳞翅目昆虫细胞的N-糖基化途径。结果表明，gp 64含有甘露糖、岩藻糖，可能还含有N-乙酰葡糖胺，但没有检测到半乳糖或唾液酸。这些相同的结果在来自草地贪夜蛾（Spodoptera frugiperda）、粉纹夜蛾（Trichoplusia ni）或Estigmene acres的三种不同鳞翅目昆虫细胞系中的任何一种中观察到，无论其是在感染后相对较早还是较晚的时间产生。这些结果表明，在AcMNPV感染的鳞翅目昆虫细胞中产生的gp 64缺乏含有外链半乳糖和唾液酸的复杂N-连接寡糖。相比之下，在哺乳动物细胞中产生的gp 64含有半乳糖和唾液酸，和内切糖苷酶diglysine显示，这些糖的N-连接，而不是O-连接，寡糖的成分。这表明gp 64上至少一个N-连接的侧链具有被加工成复合物形式的潜力。总之，这些结果表明，AcMNPV感染的鳞翅目昆虫细胞不能将gp 64上的任何N-连接的侧链转化为复杂的结构，或者外链半乳糖和唾液酸残基被添加到gp 64上，然后通过细胞或病毒外切糖苷酶除去。(C)出版社：Academic Press
The baculovirus-insect cell system is used routinely for foreign glycoprotein production, but the precise nature of the N-glycosylation pathway in this system remains unclear. Some studies indicate that these cells cannot process N-linked oligosaccharides to complex forms containing outer-chain galactose and sialic acid, while others indicate that they can, In this study, we used the major virion envelope glycoprotein of the baculovirus Autographa californica multicapsid nuclear polyhedrosis virus (AcMNPV) to probe the N-glycosylation pathway in baculovirus-infected lepidopteran insect cells. The results showed that gp64 contained mannose, fucose, and probably N-acetylglucosamine, but no detectable galactose or sialic acid, These same results were observed with gp64 produced in any one of three different lepidopteran insect cell lines derived from Spodoptera frugiperda, Trichoplusia ni, or Estigmene acres, whether it was produced at relatively earlier or later times after infection. These results indicated that the gp64 produced in AcMNPV-infected lepidopteran insect cells lacks complex N-linked oligosaccharides containing outer-chain galactose and sialic acid. By contrast, gp64 produced in mammalian cells contained both galactose and sialic acid, and endoglycosidase digestions revealed that these sugars were constituents of N-linked, not O-linked, oligosaccharides. This showed that al least one N-linked side chain on gp64 has the potential to be processed to a complex form. Together, these results suggest either that AcMNPV-infected lepidopteran insect cells are unable to convert any of the N-linked side chains on gp64 to complex structures or that outer-chain galactose and sialic acid residues are added to gp64 and then removed by cellular or viral exoglycosidases. (C) 1995 Academic Press, Inc.