N-linked glycans of Bombyx mori nucleopolyhedrovirus fibroblast growth factor are crucial for its secretion
N-linked glycans of Bombyx mori nucleopolyhedrovirus fibroblast growth factor are crucial for its secretion
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DOI：
10.1016/j.bbrc.2006.10.001
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发表时间：
2006-12-01
影响因子：
3.1
通讯作者：
Shimada, Toru
中科院分区：
文献类型：
--
作者：
Katsuma, Susumu;Daimon, Takaaki;Shimada, Toru
家蚕核多角体病毒 (BmNPV) 成纤维细胞生长因子 (BmFGF) 是一种糖基化蛋白，可有效分泌到培养基中。在这里，我们构建了表达 His 标记的野生型 (wt) 或突变 BmFGF 的突变 NPV，并表明天冬酰胺 44 和 171 这两个残基是 BmFGF 的糖基化位点。此外，从 BmFGF 中去除 N 连接聚糖导致几乎完全抑制分泌到培养基中，这表明 BmFGF 的 N 连接聚糖是其分泌所必需的。这些残基在密切相关的苜蓿银纹夜蛾 NPV (AcMNPV) 编码的 vFGF (AcFGF) 中并不保守。蛋白质印迹分析表明 AcFGF 未糖基化且分泌不良。具有两个N-连接糖基化位点的突变AcFGF比野生型AcFGF更丰富地分泌到培养基中。这是第一个直接证据表明N-连接聚糖在杆状病毒蛋白的分泌过程中的作用。 (c) 2006 Elsevier Inc. 保留所有权利。
Bombyx mori nucleopolyhedrovirus (BmNPV) fibroblast growth factor (BmFGF) is a glycosylated protein that is efficiently secreted into the medium. Here, we constructed mutant NPVs expressing His-tagged wild-type (wt) or mutant BmFGFs and showed that the two residues, asparagine 44 and 171, are the glycosylation sites of BmFGF. Also, removal of N-linked glycans from BmFGF resulted in almost complete inhibition of the secretion into the medium, suggesting that N-linked glycans of BmFGF are required for its secretion. These residues are not conserved in closely related Autographa californica NPV (AcMNPV)-encoded vFGF (AcFGF). Western blot analysis suggested that AcFGF is not glycosylated and is poorly secreted. A mutant AcFGF possessing two N-linked glycosylation sites was secreted into the medium more abundantly than that which occurred for wt AcFGF. This is the first direct evidence showing the role of N-linked glycans in the secretion process of a baculovirus protein. (c) 2006 Elsevier Inc. All rights reserved.