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首页> 外文期刊>Proceedings of the National Academy of Sciences of the United States of America >The 3-dimensional structure of a hepatitis C virus p7 ion channel by electron microscopy
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The 3-dimensional structure of a hepatitis C virus p7 ion channel by electron microscopy

机译：丙型肝炎病毒p7离子通道的3维结构电子显微镜

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摘要

Infection with the hepatitis C virus (HCV) has a huge impact on global health putting more than 170 million people at risk of developing severe liver disease. The HCV encoded p7 ion channel is essential for the production of infectious viruses. Despite a growing body of functional data, little is known about the 3-dimensional (3D) structure of the channel. Here, we present the 3D structure of a full-length viroporin, the detergent-solubilized hex-americ 42 kDa form of the HCV p7 ion channel, as determined by single-particle electron microscopy using the random conical tilting approach. The reconstruction of such a small protein complex was made possible by a combination of high-contrast staining, the symmetry, and the distinct structural features of the channel. The orientation of the p7 monomers within the density was established using immunolabeling with N and C termini specific F_(ab) fragments. The density map at a resolution of ≈16 A reveals a flower-shaped protein architecture with protruding petals oriented toward the ER lumen. This broadest part of the channel presents a comparatively large surface area providing potential interaction sites for cellular and virally encoded ER resident proteins.
机译：丙型肝炎病毒（HCV）感染对全球健康产生巨大影响，使超过1.7亿人处于发展为严重肝病的危险中。 HCV编码的p7离子通道对于生产感染性病毒至关重要。尽管功能数据越来越多，但对通道的3维（3D）结构知之甚少。在这里，我们介绍了全长维罗帕林的3D结构，HCV p7离子通道的去污剂可溶解的六聚体42 kDa形式，通过使用随机锥形倾斜方法的单粒子电子显微镜确定。通过结合高对比度染色，对称性和通道的独特结构特征，可以重建这种小蛋白质复合物。使用N和C末端特异性F_（ab）片段进行免疫标记，确定密度内p7单体的方向。分辨率约为≈16 A的密度图揭示了一种花形蛋白质结构，其突出的花瓣朝向ER内腔。通道的最宽部分呈现出较大的表面积，为细胞和病毒编码的ER驻留蛋白提供了潜在的相互作用位点。

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  • 作者

    Philipp Luik; Chee Chew; Jussi Aittoniemi; Jason Chang; Paul Wentworth. Jr; Raymond A. Dwek; Philip C. Biggin; Catherine Venien-Bryan; Nicole Zitzmann;

  • 作者单位

    Department of Biochemistry and Oxford Glycobiology Institute, University of Oxford, South Parks Road, Oxford OX1 3QU, United Kingdom;

    Department of Biochemistry and Structural Bioinformatics and Computational Biochemistry, University of Oxford, South Parks Road, Oxford OX1 3QU, United Kingdom;

    Department of Biochemistry and Structural Bioinformatics and Computational Biochemistry, University of Oxford, South Parks Road, Oxford OX1 3QU, United Kingdom;

    Department of Biochemistry and The Scripps/Oxford Laboratory, University of Oxford, South Parks Road, Oxford OX1 3QU, United Kingdom;

    Department of Biochemistry and The Scripps/Oxford Laboratory, University of Oxford, South Parks Road, Oxford OX1 3QU, United Kingdom;

    Department of Biochemistry and Oxford Glycobiology Institute, University of Oxford, South Parks Road, Oxford OX1 3QU, United Kingdom;

    Department of Biochemistry and Structural Bioinformatics and Computational Biochemistry, University of Oxford, South Parks Road, Oxford OX1 3QU, United Kingdom;

    Department of Biochemistry and Laboratory of Molecular Biophysics, University of Oxford, South Parks Road, Oxford OX1 3QU, United Kingdom;

    Department of Biochemistry and Oxford Glycobiology Institute, University of Oxford, South Parks Road, Oxford OX1 3QU, United Kingdom;

  • 收录信息 美国《科学引文索引》(SCI);美国《生物学医学文摘》(MEDLINE);美国《化学文摘》(CA);
  • 原文格式 PDF
  • 正文语种 eng
  • 中图分类
  • 关键词

    membrane protein; viroporin; single particle analysis; random conical tilt reconstruction;

    机译：膜蛋白维罗帕林单颗粒分析;随机锥形倾斜重建;

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