用单粒子方法观察规范性和药物诱导的病毒自组装之间的竞

StephenW

J Am Chem Soc. 2018 Dec 11. doi: 10.1021/jacs.8b10131. [Epub ahead of print]
Competition between Normative and Drug-Induced Virus Self-Assembly Observed with Single-Particle Methods.
Kondylis P, Schlicksup CJ, Brunk NE, Zhou J, Zlotnick A, Jacobson SC.
Abstract

Disruption of virus capsid assembly has compelling antiviral potential that has been applied to Hepatitis B Virus (HBV). HBV core protein assembly can be modulated by heteroaryldihydropyrimidines (HAPs), such molecules are collectively termed core protein allosteric modulators (CpAMs). Though the antiviral effects of CpAMs are acknowledged, the mechanism of action remains an open question. Challenging aspects of characterizing misdirected assembly are the large size and non-uniform nature of the final particles. In this study of HBV assembly, we observed a competition between normative and CpAM-induced aberrant assembly with electron microscopy and single particle nanofluidic techniques. This competition was a function of the strength of the association energy between individual core proteins, which is proportional to ionic strength. With strong association energy, assembly reactions primarily yielded morphologically normal HBV capsids, despite the presence of HAP. At weak association energy, HAPs led to increased assembly product size and disrupted morphology. The smallest particles were T = 4 icosahedra, whereas the larger particles were defective spheres, ellipsoids, and bacilliform cylinders, with regions of T = 4 geometry interspersed with flat regions. Deviation from the spherical T = 4 geometry progressively increased with particle size, which is consistent with the interpretation of a competition between two alternative assembly pathways.

PMID:
    30537810
DOI:
    10.1021/jacs.8b10131

StephenW

J Am Chem Soc。 2018年12月11日doi：10.1021 / jacs.8b10131。 [提前打印]
用单粒子方法观察规范性和药物诱导的病毒自组装之间的竞争。
Kondylis P，Schlicksup CJ，Brunk NE，Zhou J，Zlotnick A，Jacobson SC。
抽象

病毒衣壳组装的破坏具有令人信服的抗病毒潜力，已应用于乙型肝炎病毒（HBV）。 HBV核心蛋白装配可以通过杂芳基二氢嘧啶（HAP）调节，这种分子统称为核心蛋白质变构调节剂（CpAM）。尽管CpAMs的抗病毒作用得到了认可，但作用机制仍然是一个悬而未决的问题。表征错误组装的挑战性方面是最终颗粒的大尺寸和不均匀性质。在这项关于HBV组装的研究中，我们观察到规范性和CpAM诱导的异常组装与电子显微镜和单粒子纳米流体技术之间的竞争。该竞争是各核心蛋白质之间的缔合能量强度的函数，其与离子强度成比例。尽管存在HAP，但由于具有强关联能量，组装反应主要产生形态正常的HBV衣壳。在弱缔合能量下，HAP导致组装产品尺寸增加和形态破坏。最小的颗粒是T = 4二十面体，而较大的颗粒是有缺陷的球体，椭圆体和杆状圆柱体，其中T = 4几何区域散布有平坦区域。球形T = 4几何形状的偏差随颗粒尺寸逐渐增加，这与两种可选组装路径之间竞争的解释一致。

结论：
    30537810
DOI：
    10.1021 / jacs.8b10131