Binding characterization of determinants in porcine aminopeptidase N, the cellular receptor for transmissible gastroenteritis virus.
Binding characterization of determinants in porcine aminopeptidase N, the cellular receptor for transmissible gastroenteritis virus.
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猪氨肽酶 N（传染性胃肠炎病毒的细胞受体）决定簇的结合特征
DOI：
10.1016/j.jbiotec.2010.07.019
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发表时间：
2010-10-01
影响因子：
4.1
通讯作者：
Liu B
中科院分区：
文献类型：
--
作者：
Ren X;Li G;Liu B
在原核细胞中表达了4个截短型猪氨肽酶N（pAPN，猪冠状病毒的细胞受体）蛋白。用酶联免疫吸附试验（ELISA）和免疫印迹法研究pAPN特异性血清对这些蛋白的识别。用ELISA法分析了该蛋白与猪传染性胃肠炎病毒（TGEV）的结合能力。利用噬斑试验分析这些蛋白对TGEV细胞感染的抑制作用。结果表明，3个截短的pAPN与特异性抗血清呈阳性反应，其主要结合区域为36 - 223 aa、349 - 591 aa和592- 963 aa。这些蛋白质对pAPN抗体或TGE病毒粒子显示出不同的结合活性。此外，截短的蛋白质在不同程度上阻断了TGEV对细胞的感染。结果表明，pAPN的主要抗体结合结构域可能与受体结合决定簇相关。APN的作用进行了讨论的上下文中的病毒受体的使用。
Four truncated porcine aminopeptidase N (pAPN, a cellular receptor for porcine coronaviruses) proteins were expressed in prokaryotic cells. The recognizing of a specific serum against pAPN to these proteins was investigated by enzyme-linked immunosorbent assay (ELISA) and immunoblotting. The binding ability of the proteins to transmissible gastroenteritis virus (TGEV), a porcine coronavirus, was analyzed by ELISA. The inhibitory effect of these proteins to cell infection by TGEV was analyzed using plaque assays. Our data indicate that three truncated pAPNs positively reacted with the specific antiserum and the major binding regions of pAPN were limited in regions 36aa–223aa, 349aa–591aa and 592–963aa. The proteins showed discrepant binding activity to either pAPN antibody or TGE virions. Moreover, the truncated proteins blocked the infection of cells by TGEV to different extent. The results suggest that the major antibody-binding domains of pAPN may associate with the receptor-binding determinants. The role of APN is discussed in the context of virus receptor usage.
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