Adeno-associated virus capsid assembly is divergent and stochastic.
Adeno-associated virus capsid assembly is divergent and stochastic.
复制标题
腺相关病毒衣壳的组装是发散的和随机的。
DOI：
10.1038/s41467-021-21935-5
复制
发表时间：
2021-03-12
影响因子：
16.6
通讯作者：
Heck AJR
中科院分区：
文献类型：
--
作者：
Wörner TP;Bennett A;Habka S;Snijder J;Friese O;Powers T;Agbandje-McKenna M;Heck AJR
腺相关病毒(aav)越来越多地被用作基因治疗载体。aav将其基因组包装在约3.8兆道尔顿的非包膜T = 1二十面体衣壳中，该衣壳由3种不同病毒蛋白（VPs）的60个亚基组成，仅在其n端变化。虽然这三种VPs都在细胞进入和转导中发挥作用，但它们在衣壳中的精确化学计量和结构组织仍然难以捉摸。本文采用高分辨率天然质谱法研究几种AAV血清型的组成。我们的数据表明，衣壳是随机组装的，导致衣壳的组成具有高度异质性，即使是最丰富的单一VP化学计量也只占AAV总种群的一小部分。我们估计，在一种特定的制剂中，几乎每一种AAV衣壳都有独特的组成。针对实验原生质谱数据的系统模拟评分提供了一种敏感的新方法来表征这些治疗上重要的异质衣壳。腺相关病毒（aav）已成为一种很有前景的基因治疗载体。AAV衣壳由三种不同的病毒蛋白（VPs）组成的60个亚基组成。作者记录了完整AAV衣壳的高分辨率天然质谱，以评估血清型中的VP化学计量学，并揭示了不同组成的衣壳的极端异质性群体。
Adeno-associated viruses (AAVs) are increasingly used as gene therapy vectors. AAVs package their genome in a non-enveloped T = 1 icosahedral capsid of ~3.8 megaDalton, consisting of 60 subunits of 3 distinct viral proteins (VPs), which vary only in their N-terminus. While all three VPs play a role in cell-entry and transduction, their precise stoichiometry and structural organization in the capsid has remained elusive. Here we investigate the composition of several AAV serotypes by high-resolution native mass spectrometry. Our data reveal that the capsids assemble stochastically, leading to a highly heterogeneous population of capsids of variable composition, whereby even the single-most abundant VP stoichiometry represents only a small percentage of the total AAV population. We estimate that virtually every AAV capsid in a particular preparation has a unique composition. The systematic scoring of the simulations against experimental native MS data offers a sensitive new method to characterize these therapeutically important heterogeneous capsids. Adeno-associated viruses (AAVs) have emerged as promising gene therapy vectors.The AAV capsid consists of 60 subunits made up from three distinct viral proteins (VPs). Here authors record high-resolution native mass spectra of intact AAV capsids to assess the VP stoichiometries in a panel of serotypes and reveals an extremely heterogeneous population of capsids of variable composition.
登录
查看更多内容
影响因子：
12.4
作者：
Kohlbrenner, E;Aslanidi, G;Zolotukhin, S
通讯作者：
Zolotukhin, S
影响因子：
8.8
作者：
Maurer, Anna C.;Pacouret, Simon;Vandenberghe, Luk H.
通讯作者：
Vandenberghe, Luk H.
影响因子：
5.4
作者：
Padron, E;Bowman, V;Agbandje-McKenna, M
通讯作者：
Agbandje-McKenna, M
影响因子：
5.4
作者：
Popa-Wagner, Ruth;Porwal, Manvi;Kleinschmidt, Juergen A.
通讯作者：
Kleinschmidt, Juergen A.
影响因子：
5.4
作者：
Kronenberg, S;Böttcher, B;Kleinschmidt, JA
通讯作者：
Kleinschmidt, JA