Disassembly of Single Virus Capsids Monitored in Real Time with Multicycle Resistive-Pulse Sensing.
Disassembly of Single Virus Capsids Monitored in Real Time with Multicycle Resistive-Pulse Sensing.
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用多路电阻脉冲传感器对真实的实时分离的单个病毒衣壳进行检测。
DOI：
10.1021/acs.analchem.1c03855
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发表时间：
2022-01-18
影响因子：
7.4
通讯作者：
Jacobson SC
中科院分区：
文献类型：
--
作者：
Zhou J;Zlotnick A;Jacobson SC
病毒组装和拆卸是病毒生命周期中的关键步骤;然而，病毒拆卸比组装更不容易理解。对于B型肝炎病毒（HBV）衣壳，在盐酸胍（GuHCl）存在下病毒衣壳的解体表现出强烈的滞后，这需要额外的化学能来启动解体并破坏衣壳结构。为了研究HBV衣壳的分解，我们将T = 4的HBV衣壳与1.0至3.0 M的盐酸胍混合，通过随机选择颗粒来监测反应随时间的变化，并用连续脉冲传感测量它们的大小。粒子前后循环多次，以增加观察时间和观察拆卸事件的可能性。用于连续脉冲感测的四孔装置在移位期间为每个颗粒产生四个电流脉冲，这改善了对单个颗粒的跟踪和识别，并在对脉冲进行平均时提高了颗粒尺寸测量的精度。我们研究了在盐酸胍浓度低于和高于二聚体，HBV衣壳装配的基本单位的变性条件下的拆卸。正如预期的那样，在低GuHCl浓度下，衣壳几乎没有显示出分解（例如，1.0 M GuHCl），而在较高的GuHCl浓度（≥ 1.5 M）下，衣壳表现出解体，有时表现为一系列复杂的事件。在所有情况下，解体都是一个加速过程，衣壳在达到临界稳定性的几百毫秒内灾难性地解体;在衣壳解体之前，解体速率达到每秒数十个二聚体。一些拆卸事件表现出亚稳态的中间体，似乎失去一个或多个三聚体的二聚体在逐步的方式。
Virus assembly and disassembly are critical steps in the virus lifecycle; however, virus disassembly is much less well understood than assembly. For hepatitis B virus (HBV) capsids, disassembly of the virus capsid in the presence of guanidine hydrochloride (GuHCl) exhibits strong hysteresis that requires additional chemical energy to initiate disassembly and disrupt the capsid structure. To study disassembly of HBV capsids, we mixed T = 4 HBV capsids with 1.0 to 3.0 M GuHCl, monitored the reaction over time by randomly selecting particles, and measured their size with resistive-pulse sensing. Particles were cycled forward and backward multiple times to increase the observation time and likelihood of observing a disassembly event. The four-pore device used for resistive-pulse sensing produces four current pulses for each particle during translocation that improves tracking and identification of single particles and increases the precision of the particle-size measurements when the pulses are averaged. We studied disassembly at GuHCl concentrations below and above denaturing conditions of the dimer, the fundamental unit of HBV capsid assembly. As expected, capsids showed little disassembly at low GuHCl concentrations (e.g., 1.0 M GuHCl), whereas at higher GuHCl concentrations (≥ 1.5 M), capsids exhibited disassembly, sometimes as a complex series of events. In all cases, disassembly was an accelerating process, where capsids catastrophically disassembled within a few 100 ms of reaching critical stability; disassembly rates reached tens of dimers per second just before capsids fell apart. Some disassembly events exhibited metastable intermediates that appeared to lose one or more trimers of dimers in a stepwise fashion.
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影响因子：
7.7
作者：
Hadden JA;Perilla JR;Schlicksup CJ;Venkatakrishnan B;Zlotnick A;Schulten K
通讯作者：
Schulten K
影响因子：
17.1
作者：
Edwards, Martin A.;German, Sean R.;White, Henry S.
通讯作者：
White, Henry S.
影响因子：
3
作者：
Kumar, Rajesh;Bhuyan, Abani K.
通讯作者：
Bhuyan, Abani K.
影响因子：
1.6
作者：
DEBLOIS, RW;BEAN, CP
通讯作者：
BEAN, CP
影响因子：
3.3
作者：
Moerman, Pepijn;van der Schoot, Paul;Kegel, Willem
通讯作者：
Kegel, Willem