Alanine scanning mutagenesis of hepatitis C virus E2 cysteine residues: Insights into E2 biogenesis and antigenicity.
Alanine scanning mutagenesis of hepatitis C virus E2 cysteine residues: Insights into E2 biogenesis and antigenicity.
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DOI：
10.1016/j.virol.2013.10.020
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发表时间：
2014-01
期刊：
影响因子：
3.7
通讯作者：
Wenbo Wang;Mo Guan;Yuan Liu;Qingqiang Xu;Haoran Peng;Xiaoqing Liu;Zi-wei Tang;Yongzhe Zhu;Da-Ge Wu;H. Ren;P. Zhao;Z. Qi
中科院分区：
文献类型：
--
作者：
Wenbo Wang;Mo Guan;Yuan Liu;Qingqiang Xu;Haoran Peng;Xiaoqing Liu;Zi-wei Tang;Yongzhe Zhu;Da-Ge Wu;H. Ren;P. Zhao;Z. Qi
丙型肝炎病毒包膜糖蛋白2（E2）的胞外域含有18个保守的半胱氨酸残基。通过半胱氨酸-丙氨酸突变和功能分析，发现H77 E2中的6个Cys（C494，C508，C552，C564，C607和C644）是构象依赖性mAb H53识别H77 E2所必需的。去除这些Cys残基中的任何一个都不影响E2与E1的异源二聚化，但显著降低了E1 E2的跨膜转运。这些Cys与C429和C503一起是构象依赖性mAb H48识别所需的。除C607外，所有上述Cys都是H77和Con 1 E2与CD 81结合所必需的。上述Cys的突变均不影响E2诱导小鼠产生中和抗体的能力。小鼠抗体主要识别E2线性表位，与人E2抗体识别的表位无关。这些发现为理解功能性HCV包膜蛋白的生物发生和HCV中和免疫提供了新的见解。
Envelope glycoprotein 2 (E2) of hepatitis C virus contains 18 conserved cysteine (Cys) residues in its ectodomain. By cysteine-alanine mutagenesis and function analysis, six Cys in H77 E2 (C494, C508, C552, C564, C607 and C644) were found to be indispensable for recognition by conformation-dependent mAb H53. Removal of any of these Cys residues did not affect E2 heterodimerization with E1, but notably reduced E1E2 transmembrane transportation. These Cys together with C429 and C503 were required for conformation-dependent mAb H48 recognition. All of the above Cys except C607 were required for H77 and Con1 E2 binding to CD81. None of individual mutation of above Cys affected the ability of E2 to induce neutralizing antibodies in mice. Mouse antibodies mainly recognize E2 linear epitopes and are unrelated to epitopes recognized by human E2 antibodies. The findings provide new insights for understanding the biogenesis of functional HCV envelope proteins and HCV neutralizing immunity.