Identification of the amino terminal subunit of the glycoprotein of Borna disease virus
Identification of the amino terminal subunit of the glycoprotein of Borna disease virus
复制标题
博尔纳病病毒糖蛋白氨基末端亚基的鉴定
DOI：
10.1016/s0014-5793(02)03513-5
复制
发表时间：
2002
期刊：
影响因子：
3.5
通讯作者：
M. Eickmann
中科院分区：
文献类型：
--
作者：
S. Kiermayer;Ina Kraus;J. Richt;W. Garten;M. Eickmann
博尔纳病病毒（Borna disease virus，BDV）是博尔纳病病毒（Borna disease virus，BDV）的唯一一种表面膜糖蛋白，其分子量为57 kDa，N-糖基化后为约94 kDa的前体糖蛋白（precursor glycoprotein，GP）。它被细胞蛋白酶弗林蛋白酶加工成C-末端膜锚定亚基GP-C，也称为gp 43，和推测的N-末端亚基GP-N，其高度糖基化，分子量约为51 kDa。然而，到目前为止，后者仍然未检测到的BDV感染的材料。我们描述了一种新的方法来确定聚糖掩蔽的线性抗原表位。在本研究中，GP-N被确定在BDV感染的细胞通过凝集素沉淀，酶促去糖基化印迹和免疫化学的组合，使用N-末端特异性抗血清。GP-N在其糖基化形式中具有45-50 kDa的表观分子量，在其去糖基化形式中具有27 kDa的表观分子量。N-聚糖分析显示，前体GP仅含有富含甘露糖的N-聚糖，而GP-N和GP-C含有富含甘露糖的复合型N-聚糖。
The only surface membrane glycoprotein of Borna disease virus (BDV) is synthesized as a polypeptide with a molecular mass of 57 kDa and N-glycosylated to a precursor glycoprotein (GP) of about 94 kDa. It is processed by the cellular protease furin into the C-terminal membrane-anchored subunit GP-C, also known as gp43, and a presumptive N-terminal subunit GP-N, that is highly glycosylated and has a molecular mass of about 51 kDa. However, up to now the latter remained undetected in BDV-infected material. We describe a novel approach to identify glycan masked linear antigenic epitopes. In the present study, GP-N was identified in BDV-infected cells by a combination of lectin precipitation, enzymatic deglycosylation on blot and immunochemistry using an N-terminal specific antiserum. The GP-N has an apparent molecular mass of 45–50 kDa in its glycosylated form and 27 kDa in its deglycosylated form. N-glycan analysis revealed that the precursor GP contains only mannose-rich N-glycans, whereas GP-N and GP-C contain mannose-rich and complex-type N-glycans.