Alterations in the structure of the oligosaccharide of vesicular stomatitis virus G protein by swainsonine.
Alterations in the structure of the oligosaccharide of vesicular stomatitis virus G protein by swainsonine.
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苦马豆素对水疱性口炎病毒 G 蛋白寡糖结构的改变。
DOI：
10.1128/jvi.46.1.60-69.1983
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发表时间：
1983
影响因子：
5.4
通讯作者：
Elbein,AD
中科院分区：
文献类型：
--
作者：
Kang,MS;Elbein,AD
苦马豆素是一种糖蛋白加工抑制剂，可抑制水泡性口炎病毒G蛋白正常寡糖链的形成。因此，当水泡性口炎病毒在苦马豆素（15 - 500 ng/ml）存在下在幼仓鼠肾细胞中生长并用[2- 3 H]甘露糖标记时，G蛋白的寡糖部分完全易受内切氨基葡萄糖苷酶H的作用。然而，正常的病毒糖蛋白对这种酶不敏感。甘露糖标记的寡糖的各种酶处理和甲基化研究表明苦马豆素引起具有中性寡糖特征的寡甘露糖基核心（Man 5GlcNAc 2-Asn）加上复杂寡糖特征的分支结构（NeuNAc-Gal-GlcNAc）的混合型寡糖的形成。提出了这种混合寡糖的结构。苦马豆素对[14 C]亮氨酸掺入病毒蛋白没有影响，也没有改变这些培养物中产生的PFU数量。然而，它确实略微降低了[3 H]葡糖胺的掺入，并增加了[3 H]甘露糖的掺入。水泡性口炎病毒提出的苦马豆素的存在下绑定更紧密的柱的伴刀豆球蛋白A-琼脂糖凝胶比对照病毒。苦马豆素必须在病毒感染的前4或5小时内加入才有效。因此，当在感染的前3小时内的任何时间每ml加入100 ng生物碱时，基本上所有的糖蛋白都易于被内切氨基葡萄糖苷酶H消化。然而，当苦马豆素加入4小时后开始感染，30%的糖肽成为耐内切氨基葡萄糖苷酶H;在5小时，70%的耐药。苦马豆素的作用是可逆的，因为除去生物碱后，细胞可以形成正常的复合糖蛋白。然而，就寡糖结构而言，去除时间是关键的。
Swainsonine, an inhibitor of glycoprotein processing, inhibits the formation of the normal oligosaccharide chain of the G protein of vesicular stomatitis virus. Thus, when vesicular stomatitis virus was grown in baby hamster kidney cells in the presence of swainsonine (15 to 500 ng/ml) and labeled with [2-3H]mannose, the oligosaccharide portion of the G protein was completely susceptible to the action of endoglucosaminidase H. However, the normal viral glycoprotein is not susceptible to this enzyme. Various enzymatic treatments and methylation studies of the mannose-labeled oligosaccharides suggest that swainsonine causes the formation of a hybrid-type oligosaccharide having an oligomannosyl core (Man5GlcNAc2-Asn) characteristic of neutral oligosaccharides plus the branch structure (NeuNAc-Gal-GlcNAc) characteristic of the complex oligosaccharides. A structure for this hybrid oligosaccharide is proposed. Swainsonine had no effect on the incorporation of [14C]leucine into viral proteins, nor did it change the number of PFU produced in these cultures. It did, however, slightly decrease the incorporation of [3H]glucosamine and increase the incorporation of [3H]mannose. Vesicular stomatitis virus raised in the presence of swainsonine bound much more tightly to columns of concanavalin A-Sepharose than did control virus. Swainsonine had to be added within the first 4 or 5 h of virus infection to be effective. Thus, when 100 ng of the alkaloid per ml was added at any time within the first 3 h of infection, essentially all of the glycoprotein was susceptible to digestion by endoglucosaminidase H. However, when swainsonine was added 4 h after the start of infection, 30% of the glycopeptides became resistant to endoglucosaminidase H; at 5 h, 70% were resistant. The effect of swainsonine was reversible since removal of the alkaloid allowed the cells to form the normal complex glycoproteins. However, the time of removal was critical in terms of oligosaccharide structure.
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