Lombana Laura, Ortega-Atienza Sara, Gómez-Gutiérrez Julián, Yélamos Belén, Peterson Darrell L, Gavilanes Francisco
Department of Biochemistry and Molecular Biology, Facultad de Ciencias Químicas, Universidad Complutense de Madrid, 28040 Madrid, Spain.
Department of Biochemistry and Molecular Biology, Medical College of Virginia, Virginia Commonwealth University, Richmond, VA 23298, USA.
Virus Res. 2016 Jun 2;217:63-70. doi: 10.1016/j.virusres.2016.02.009. Epub 2016 Mar 2.
We have obtained a chimeric protein containing the ectodomains of hepatitis C virus (HCV) envelope proteins but lacking the region 268-292 of E1. All its structural properties are coincident with those of the corresponding full length chimera. The deleted and entire chimeras were compared in terms of their membrane destabilizing properties. No differences were found in their ability to induce vesicle aggregation and lipid mixing but the deleted chimera showed a reduced capacity to promote leakage. The role of the deletion was also studied by obtaining HCV pseudoparticles (HCVpp). Both E1 and E2, and also the E1 deleted mutant, were incorporated into HCVpp to a similar level. However, HCVpp containing the E1 deleted protein are almost unable to infect Huh7 cells. These results point to the involvement of the region 268-292 in the formation of pores in the membrane necessary for the complete fusion of the membranes.
我们获得了一种嵌合蛋白，其包含丙型肝炎病毒（HCV）包膜蛋白的胞外结构域，但缺少E1的268 - 292区域。其所有结构特性均与相应全长嵌合体的结构特性一致。对缺失型和完整型嵌合体的膜去稳定特性进行了比较。在诱导囊泡聚集和脂质混合的能力方面未发现差异，但缺失型嵌合体促进渗漏的能力降低。还通过制备HCV假颗粒（HCVpp）研究了缺失区域的作用。E1和E2以及E1缺失突变体均以相似水平掺入HCVpp中。然而，含有E1缺失蛋白的HCVpp几乎无法感染Huh7细胞。这些结果表明268 - 292区域参与了膜融合所需的膜孔形成。
