Formation of native hepatitis C virus glycoprotein complexes
Formation of native hepatitis C virus glycoprotein complexes
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DOI：
10.1128/jvi.71.1.697-704.1997
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发表时间：
1997-01-01
影响因子：
5.4
通讯作者：
Dubuisson, J
中科院分区：
文献类型：
--
作者：
Deleersnyder, V;Pillez, A;Dubuisson, J
丙型肝炎病毒（HCV）糖蛋白（E1和E2）相互作用形成异二聚体复合物，该复合物已被认为是HCV病毒粒子包膜的功能亚基。在细胞培养瞬态表达实验中，正确折叠、非共价相关E1E2复合物的形成是一个缓慢而低效的过程。由于缺乏适当的免疫试剂，很难区分经过生产性折叠和组装的糖蛋白分子与那些遵循非生产性途径导致错误折叠和聚集的糖蛋白分子。在这里，我们报道了一个构象敏感e2反应性单克隆抗体（H2）的分离和鉴定。H2单克隆抗体选择性识别缓慢成熟的E1E2异源二聚体，这些异源二聚体是非共价连接的，具有蛋白酶抗性，并且不再与内质网伴侣钙连联蛋白相关。该复合物可能代表了HCV糖蛋白异二聚体的原生芽前形式。除了为HCV病毒粒子组装和进入的基础研究提供一种新的试剂外，该单克隆抗体还可用于优化HCV糖蛋白复合物的生产和分离，用于血清诊断和疫苗应用。
The hepatitis C virus (HCV) glycoproteins (E1 and E2) interact to form a heterodimeric complex, which has been proposed as a functional subunit of the HCV virion envelope. As examined in cell culture transient-expression assays, the formation of properly folded, noncovalently associated E1E2 complexes is a slow and inefficient process. Due to lack of appropriate immunological reagents, it has been difficult to distinguish between glycoprotein molecules that undergo productive folding and assembly from those which follow a nonproductive pathway leading to misfolding and aggregation. Here we report the isolation and characterization of a conformation-sensitive E2-reactive monoclonal antibody (H2). The H2 monoclonal antibody selectively recognizes slowly maturing E1E2 heterodimers which are noncovalently linked, protease resistant, and no longer associated with the endoplasmic reticulum chaperone calnexin. This complex probably represents the native prebudding form of the HCV glycoprotein heterodimer. Besides providing a novel reagent for basic studies on HCV virion assembly and entry, this monoclonal antibody should be useful for optimizing production and isolation of native HCV glycoprotein complexes for serodiagnostic and vaccine applications.