乙型肝炎病毒衣壳蛋白二聚体的动力学通过变构网络调节装

StephenW

Dynamics of Hepatitis B virus capsid protein dimer regulate assembly through an allosteric network
Angela Patterson, Zhongchao Zhao, Elizabeth Waymire, Adam Zlotnick, Brian Bothner

    PMID: 32662972 DOI: 10.1021/acschembio.0c00481

Abstract

While there is an effective vaccine for Human Hepatitis B Virus (HBV), 257 million people have chronic infections for which there is no cure. The assembly process for the viral capsid is a potential therapeutic target. In order to understand the capsid assembly process, we investigated the dimeric building blocks of the capsid. To understand what blocks assembly, we took advantage of an assembly incompetent mutant dimer, Cp149-Y132A, located in the inter-dimer interface. This mutation leads to changes in protein dynamics throughout the structure of the dimer as measured by hydrogen deuterium exchange mass spectrometry (HDX-MS). To further understand how the HBV capsid assembles, the homolog woodchuck HBV (WHV) capsid protein dimer (Cp) was used. WHV is more stable than HBV in HDX-MS and native mass spectrometry experiments. Because the WHV Cp assembles more rapidly into viral capsids than HBV, it was suspected that an increase in stability of the intra-dimer interface and/or in the contact region leads to increased assembly rates. The differences in dynamics when comparing HBV and human Cp149-Y132A as well as the differences in dynamics when comparing the HBV and WHV Cps allowed us to map an allosteric network within the HBV dimer. Through a careful comparison of structure, stability and dynamics using four different capsid protein dimers, we conclude that protein subunit dynamics regulate HBV capsid assembly.

StephenW

乙型肝炎病毒衣壳蛋白二聚体的动力学通过变构网络调节装配
安吉拉·帕特森，赵忠超，伊丽莎白·韦米尔，亚当·兹洛特尼克，布莱恩·博特纳

    PMID：32662972 DOI：10.1021 / acschembio.0c00481

抽象

尽管有一种有效的人类乙型肝炎病毒（HBV）疫苗，但有2.57亿人患有无法治愈的慢性感染。病毒衣壳的组装过程是潜在的治疗目标。为了了解衣壳的组装过程，我们研究了衣壳的二聚体构建基块。要了解什么会阻止装配，我们利用了位于二聚物间界面中的装配不当的突变体二聚体Cp149-Y132A。通过氢氘交换质谱法（HDX-MS）测量，此突变导致整个二聚体结构中的蛋白质动力学变化。为了进一步了解HBV衣壳如何组装，使用了同源的土拨鼠HBV（WHV）衣壳蛋白二聚体（Cp）。在HDX-MS和天然质谱实验中，WHV比HBV更稳定。因为WHV Cp比HBV更快速地装配到病毒衣壳中，所以怀疑二聚体内界面和/或接触区域稳定性的提高会导致装配速度的提高。比较HBV和人Cp149-Y132A时的动力学差异以及比较HBV和WHV Cps时的动力学差异使我们能够绘制HBV二聚体内的变构网络。通过使用四个不同的衣壳蛋白二聚体仔细比较结构，稳定性和动力学，我们得出结论，蛋白亚基动力学调节着HBV衣壳装配。