3D variability analysis: Resolving continuous flexibility and discrete heterogeneity from single particle cryo-EM
3D variability analysis: Resolving continuous flexibility and discrete heterogeneity from single particle cryo-EM
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DOI：
10.1016/j.jsb.2021.107702
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发表时间：
2021-03-06
影响因子：
3
通讯作者：
Fleet, David J.
中科院分区：
文献类型：
--
作者：
Punjani, Ali;Fleet, David J.
单粒子低温电子显微镜在确定蛋白质分子的静态结构方面具有优势，但现有的3D重建方法在模拟柔性蛋白质方面一直效率低下。我们介绍了3D变异性分析(3DVA)，这是一种以高分辨率将构象变化的线性子空间模型拟合到低温电磁数据的算法。3DVA能够分辨和可视化大小蛋白质的详细分子运动，从单颗粒冷冻-EM数据中揭示出新的生物学见解。实验结果表明，3DVA能够分辨GPCR复合体亚50 kDa跨膜区α-螺旋的多种柔性运动、钠离子通道的弯曲模式、蛋白酶体中五种对称和对称破坏的柔性、剪接体复合体中的大运动以及核糖体组装的离散构象状态。3DVA在CryoSPARC软件包中实现。
Single particle cryo-EM excels in determining static structures of protein molecules, but existing 3D reconstruction methods have been ineffective in modelling flexible proteins. We introduce 3D variability analysis (3DVA), an algorithm that fits a linear subspace model of conformational change to cryo-EM data at high resolution. 3DVA enables the resolution and visualization of detailed molecular motions of both large and small proteins, revealing new biological insight from single particle cryo-EM data. Experimental results demonstrate the ability of 3DVA to resolve multiple flexible motions of a-helices in the sub-50 kDa transmembrane domain of a GPCR complex, bending modes of a sodium ion channel, five types of symmetric and symmetry-breaking flexibility in a proteasome, large motions in a spliceosome complex, and discrete conformational states of a ribosome assembly. 3DVA is implemented in the cryoSPARC software package.