TGEV corona virus ORF4 encodes a membrane protein that is incorporated into virions.
TGEV corona virus ORF4 encodes a membrane protein that is incorporated into virions.
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DOI：
10.1016/0042-6822(92)90521-p
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发表时间：
1992-06
期刊：
影响因子：
3.7
通讯作者：
Laude H
中科院分区：
文献类型：
--
作者：
Godet M;L'Haridon R;Vautherot JF;Laude H
通过使用杆状病毒载体表达，已证实传染性胃肠炎病毒（TGEV）开放阅读框ORF4（82个氨基酸）的编码潜能。针对10K重组产物产生的5种单克隆抗体（MAb）在TGEV感染的细胞中免疫沉淀出大小相似的多肽。在昆虫细胞和哺乳动物细胞上进行的免疫荧光分析显示，ORF4是一种膜相关蛋白，这一发现与ORF4序列中跨膜区段的预测相符。通过肽段扫描以及对截短的ORF4重组蛋白反应性的分析，确定两个表位位于该序列的最后21个C末端残基内。由于发现相关单克隆抗体可诱导细胞表面荧光，这些数据表明ORF4可能是一种具有C外向 - N内向取向的整合膜蛋白。抗ORF4单克隆抗体还用于表明在TGEV病毒体制备物中可检测到ORF4多肽，估计每个病毒粒子掺入20个分子。氨基酸序列数据的比较提供了有力证据，表明其他冠状病毒编码一种与TGEV ORF4同源的多肽。我们的研究结果使我们提出，ORF4代表一种新的次要结构多肽，暂定为SM（小膜蛋白）。
The coding potential of the open reading frame ORF4 (82 amino acids) of transmissible gastroenteritis virus (TGEV) has been confirmed by expression using a baculovirus vector. Five monoclonal antibodies (MAbs) raised against the 10K recombinant product immunoprecipitated a polypeptide of a similar size in TGEV-infected cells. Immunofluorescence assays performed both on insect and mammalian cells revealed that ORF4 was a membrane-associated protein, a finding consistent with the prediction of a membrane-spanning segment in ORF4 sequence. Two epitopes were localized within the last 21 C-terminal residues of the sequence through peptide scanning and analysis of the reactivity of a truncated ORF4 recombinant protein. Since the relevant MAbs were found to induce a cell surface fluorescence, these data suggest that ORF4 may be an integral membrane protein having a Cexo-Nendo orientation. Anti-ORF4 MAbs were also used to show that ORF4 polypeptide may be detected in TGEV virion preparations, with an estimated number of 20 molecules incorporated per particle. Comparison of amino acid sequence data provided strong evidence that other coronaviruses encode a polypeptide homologous to TGEV ORF4. Our results led us to propose that ORF4 represents a novel minor structural polypeptide, tentatively designated SM (small membrane protein).
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