Interactions of bovine viral diarrhoea virus glycoprotein Erns with cell surface glycosaminoglycans
Interactions of bovine viral diarrhoea virus glycoprotein Erns with cell surface glycosaminoglycans
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DOI：
10.1099/0022-1317-81-2-451
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发表时间：
2000-02-01
影响因子：
3.8
通讯作者：
McCauley, JW
中科院分区：
文献类型：
--
作者：
Iqbal, M;Flick-Smith, H;McCauley, JW
牛病毒性腹泻病毒 (BVDV) 的重组 E-rns 糖蛋白已标记有标记表位或连接到免疫球蛋白 Fc 尾部，并在昆虫和哺乳动物细胞系中表达。该产品被证明具有功能性，既具有核糖核酸酶活性，又能与允许和不允许 BVDV 复制的各种细胞结合。向培养基中添加可溶性 E-rns 可阻断 BVDV 在许可细胞中的复制。当用肝素酶 I 或 III 处理细胞时，表位标记的 E-rns 与许可型小牛睾丸 (CTe) 细胞的结合被消除，病毒感染减少。 E-rns 无法与缺乏糖胺聚糖（pgsA-745 细胞）或硫酸乙酰肝素（pgsD-677 细胞）的突变中国仓鼠卵巢（CHO）细胞结合，但与正常 CHO 细胞结合。 E-rns 还与固定在琼脂糖上的肝素结合，并且可以被肝素和高浓度的盐洗脱。 E-rns与CTe细胞培养物结合的流式细胞术分析表明，糖胺聚糖如肝素、岩藻依聚糖和硫酸皮肤素均抑制结合，但硫酸葡聚糖、硫酸角质素、硫酸软骨素和甘露聚糖不能抑制结合。低分子量多磺化抑制剂苏拉明也抑制与 CTe 细胞的结合，但聚-L-赖氨酸则不然。此外，苏拉明、苏拉明类似物CPD14、岩藻依聚糖和多硫酸戊聚糖抑制病毒的感染性。据推测，E-rns 与细胞的结合是通过与糖胺聚糖的相互作用而实现的，并且 BVDV 可能最初通过这种相互作用与细胞结合。
Recombinant E-rns glycoprotein of bovine viral diarrhoea virus (BVDV) has been tagged with a marker epitope or linked to an immunoglobulin Fc tail and expressed in insect and mammalian cell lines. The product was shown to be functional, both having ribonuclease activity and binding to a variety of cells that were permissive and non-permissive for replication of BVDV. Addition of soluble E-rns to the medium blocked replication of BVDV in permissive cells. Binding of epitope-tagged E-rns to permissive calf testes (CTe) cells was abolished and virus infection was reduced when cells were treated with heparinases I or III. E-rns failed to bind to mutant Chinese hamster ovary (CHO) cells that lacked glycosaminoglycans (pgsA-745 cells) or heparan sulphate (pgsD-677 cells) but bound to normal CHO cells. E-rns also bound to heparin immobilized on agarose and could be eluted by heparin and by a high concentration of salt. Flow cytometric analysis of E-rns binding to CTe cell cultures showed that glycosaminoglycans such as heparin, fucoidan and dermatan sulphate all inhibit binding but dextran sulphate, keratan sulphate, chondroitin sulphate and mannan fail to inhibit binding. The low molecular mass polysulphonated inhibitor suramin also inhibited binding to CTe cells but poly-L-lysine did not. Furthermore, suramin, the suramin analogue CPD14, fucoidan and pentosan polysulphate inhibited the infectivity of virus. It is proposed that binding of E-rns to cells is through an interaction with glycosaminoglycans and that BVDV may bind to cells initially through this interaction.