基于苯基丙烯酰胺的抗病毒药物在乙型肝炎病毒衣壳组装过

StephenW

ACS Infect Dis. 2019 Jan 7. doi: 10.1021/acsinfecdis.8b00290. [Epub ahead of print]
Evolution of Intermediates during Capsid Assembly of Hepatitis B Virus with Phenylpropenamide-based Antivirals.
Kondylis P, Schlicksup CJ, Katen SP, Lee LS, Zlotnick A, Jacobson SC.
Abstract

The self-assembly of virus capsids is a potential target for antivirals due to its importance in the virus lifecycle. Here, we investigate the effect of phenylpropenamide derivatives B-21 and AT-130 on the assembly of hepatitis B virus (HBV) core protein. Phenylpropenamides are widely believed to yield assembly of spherical particles resembling native, empty HBV capsids. Because the details of assembly can be overlooked with ensemble measurements, we performed resistive-pulse measurements on nanofluidic devices with four pores in series to characterize the size distributions of the products in real time. With its single particle sensitivity and compatibility with typical assembly buffers, resistive-pulse sensing is well suited for analyzing virus assembly in vitro. We observed that assembly with B-21 and AT-130 produced a large fraction of partially complete virus particles that may be on-path, off-path, or trapped. For both B-21 and AT-130, capsid assembly was more sensitive to disruption under conditions where the inter-protein association energy was low at lower salt concentrations. Dilution of the reaction solutions led to the rearrangement of the incomplete particles and demonstrated that these large intermediates may be on-path, but are labile, and exist in a frustrated dynamic equilibrium. During capsid assembly, phenylpropenamide molecules modestly increase the association energy of dimers, prevent intermediates from dissociating, and lead to kinetic trapping where the formation of too many capsids has been initiated leading to both empty and incomplete particles.

PMID:
    30616343
DOI:
    10.1021/acsinfecdis.8b00290

StephenW

ACS Infect Dis。 2019年1月7日doi：10.1021 / acsinfecdis.8b00290。 [提前打印]
基于苯基丙烯酰胺的抗病毒药物在乙型肝炎病毒衣壳组装过程中中间体的演变。
Kondylis P，Schlicksup CJ，Katen SP，Lee LS，Zlotnick A，Jacobson SC。
抽象

病毒衣壳的自组装由于其在病毒生命周期中的重要性而成为抗病毒药物的潜在靶标。在这里，我们研究了苯丙烯酰胺衍生物B-21和AT-130对乙型肝炎病毒（HBV）核心蛋白装配的影响。人们普遍认为苯基丙烯酰胺可以产生类似于天然空HBV衣壳的球形颗粒的组装。由于集合测量可以忽略装配细节，我们对具有四个串联孔的纳米流体装置进行电阻脉冲测量，以实时表征产品的尺寸分布。凭借其单粒子灵敏度和与典型组装缓冲液的兼容性，电阻脉冲传感非常适合分析体外病毒组装。我们观察到与B-21和AT-130的组装产生了大部分部分完整的病毒颗粒，这些颗粒可能在路径上，偏离路径或被捕获。对于B-21和AT-130，衣壳组装在较低盐浓度下蛋白质间缔合能量较低的条件下对破坏更敏感。反应溶液的稀释导致不完全颗粒的重排，并证明这些大的中间体可能在路径上，但是不稳定，并且存在于受抑制的动态平衡中。在衣壳组装期间，苯基丙烯酰胺分子适度地增加二聚体的缔合能量，防止中间体离解，并导致动力学捕获，其中已经开始形成太多衣壳，导致空的和不完整的颗粒。

结论：
    30616343
DOI：
    10.1021 / acsinfecdis.8b00290