Computational Prediction of the Heterodimeric and Higher-Order Structure of gpE1/gpE2 Envelope Glycoproteins Encoded by Hepatitis C Virus
Computational Prediction of the Heterodimeric and Higher-Order Structure of gpE1/gpE2 Envelope Glycoproteins Encoded by Hepatitis C Virus
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DOI：
10.1128/jvi.02309-16
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发表时间：
2017-04-01
影响因子：
5.4
通讯作者：
Houghton, Michael
中科院分区：
文献类型：
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作者：
Freedman, Holly;Logan, Michael R.;Houghton, Michael
尽管最近新开发的直接作用抗病毒药物对丙型肝炎取得了成功，但由于大多数携带者缺乏诊断和治疗费用高昂，该疾病仍然是全球健康威胁。丙型肝炎病毒（HCV）脂膜糖蛋白E1和E2形成的异源二聚体是一种潜在的疫苗候选物和抗病毒靶点。虽然E1/E2的结构尚未解析，但已经确定了E1和E2胞外域的部分晶体结构。结构的未解决的部分在可以用当前计算建模工具建模的范围内。此外，各种额外的实验数据可用于支持E1/E2结构的计算预测，例如来自抗体结合研究、冷冻电子显微镜（cryo-EM）、突变分析、肽结合分析、接头扫描诱变和核磁共振（NMR）研究的数据。根据这些丰富的实验数据，我们建立了全长E1/E2异二聚体的计算机模型。我们的模型支持E1/E2组装成三聚体，这是Falson和同事先前的研究提出的（P. Falson，B. Bartosch，K. Alsaleh，B. A. Tews，A. Loquet，Y.奇佐拉湖里瓦角，澳-地蒙蒂尼角蒙彼利埃湾Duverlie，E. I. Pecheur，M. le Maire，F. L. Cosset，J. Dubuisson，and F. Penin，J. Virol. 89：10333-10346，2015，https：//doi.org/10.1128/JVI.00991-15）。通过使用纯化的重组E1/E2获得的尺寸排阻色谱和Western印迹数据支持我们的假设。我们的模型表明，在病毒组装过程中，E1/E2的三聚体可以进一步组装成一个五聚体，12个五聚体组成一个单一的HCV病毒粒子。我们预计，这种新的模式将提供一个有用的框架，HCV包膜结构和抗病毒strategies.IMPORTANCE的发展一亿五千万人已被估计感染丙型肝炎病毒，更多的是在感染的风险。更好地了解负责附着和融合的HCV包膜的结构，可以帮助开发这种疾病的疫苗和/或新的治疗方法。我们利用计算技术来预测的E1/E2异源二聚体的基础上的包膜糖蛋白E1和E2的部分晶体结构的全长模型。E1/E2已经被广泛的实验研究，这提供了有价值的数据，这有助于我们在我们的建模。我们提出的结构是用来建议组织的HCV包膜。我们还提出了新的实验数据，从尺寸排阻色谱，支持我们的计算预测的E1/E2的三聚体低聚状态。
Despite the recent success of newly developed direct-acting antivirals against hepatitis C, the disease continues to be a global health threat due to the lack of diagnosis of most carriers and the high cost of treatment. The heterodimer formed by glycoproteins E1 and E2 within the hepatitis C virus (HCV) lipid envelope is a potential vaccine candidate and antiviral target. While the structure of E1/E2 has not yet been resolved, partial crystal structures of the E1 and E2 ectodomains have been determined. The unresolved parts of the structure are within the realm of what can be modeled with current computational modeling tools. Furthermore, a variety of additional experimental data is available to support computational predictions of E1/E2 structure, such as data from antibody binding studies, cryo-electron microscopy (cryo-EM), mutational analyses, peptide binding analysis, linker-scanning mutagenesis, and nuclear magnetic resonance (NMR) studies. In accordance with these rich experimental data, we have built an in silico model of the full-length E1/E2 heterodimer. Our model supports that E1/E2 assembles into a trimer, which was previously suggested from a study by Falson and coworkers (P. Falson, B. Bartosch, K. Alsaleh, B. A. Tews, A. Loquet, Y. Ciczora, L. Riva, C. Montigny, C. Montpellier, G. Duverlie, E. I. Pecheur, M. le Maire, F. L. Cosset, J. Dubuisson, and F. Penin, J. Virol. 89: 10333-10346, 2015, https://doi.org/10.1128/JVI.00991-15). Size exclusion chromatography and Western blotting data obtained by using purified recombinant E1/E2 support our hypothesis. Our model suggests that during virus assembly, the trimer of E1/E2 may be further assembled into a pentamer, with 12 pentamers comprising a single HCV virion. We anticipate that this new model will provide a useful framework for HCV envelope structure and the development of antiviral strategies.IMPORTANCE One hundred fifty million people have been estimated to be infected with hepatitis C virus, and many more are at risk for infection. A better understanding of the structure of the HCV envelope, which is responsible for attachment and fusion, could aid in the development of a vaccine and/or new treatments for this disease. We draw upon computational techniques to predict a full-length model of the E1/E2 heterodimer based on the partial crystal structures of the envelope glycoproteins E1 and E2. E1/E2 has been widely studied experimentally, and this provides valuable data, which has assisted us in our modeling. Our proposed structure is used to suggest the organization of the HCV envelope. We also present new experimental data from size exclusion chromatography that support our computational prediction of a trimeric oligomeric state of E1/E2.