Subcellular location and topology of severe acute respiratory syndrome coronavirus envelope protein.
Subcellular location and topology of severe acute respiratory syndrome coronavirus envelope protein.
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DOI：
10.1016/j.virol.2011.03.029
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发表时间：
2011-07-05
期刊：
影响因子：
3.7
通讯作者：
Enjuanes L
中科院分区：
文献类型：
--
作者：
Nieto-Torres JL;Dediego ML;Alvarez E;Jiménez-Guardeño JM;Regla-Nava JA;Llorente M;Kremer L;Shuo S;Enjuanes L
严重急性呼吸综合征（SARS）冠状病毒（CoV）包膜蛋白（E蛋白）是一种跨膜蛋白。E蛋白的几种亚细胞定位和拓扑构象已被提出。为了确定正确的，多克隆和单克隆抗体的氨基或羧基末端的E蛋白，分别产生。E蛋白主要存在于内质网-高尔基体中间区（ERGIC）中。免疫荧光、免疫电镜和细胞表面蛋白标记均未发现E蛋白存在于质膜上。此外，全细胞膜片钳测量质膜电压门控离子通道活性表明，E蛋白不存在于质膜。提出了SARS-CoV E蛋白氨基端朝向细胞内膜内腔、羧基端朝向细胞质的拓扑构象。
Severe acute respiratory syndrome (SARS) coronavirus (CoV) envelope (E) protein is a transmembrane protein. Several subcellular locations and topological conformations of E protein have been proposed. To identify the correct ones, polyclonal and monoclonal antibodies specific for the amino or the carboxy terminus of E protein, respectively, were generated. E protein was mainly found in the endoplasmic reticulum-Golgi intermediate compartment (ERGIC) of cells transfected with a plasmid encoding E protein or infected with SARS-CoV. No evidence of E protein presence in the plasma membrane was found by using immunofluorescence, immunoelectron microscopy and cell surface protein labeling. In addition, measurement of plasma membrane voltage gated ion channel activity by whole-cell patch clamp suggested that E protein was not present in the plasma membrane. A topological conformation in which SARS-CoV E protein amino terminus is oriented towards the lumen of intracellular membranes and carboxy terminus faces cell cytoplasm is proposed.