We describe a novel approach to sorting class averages of a structure in multiple conformational states in order to generate 3D reconstructions that account for conformational variability present in the sample. The method assumes that the relative Euler angles between class averages are known, then uses a common lines approach to match any given class against a set of distinct conformations from a selected view of the structure. We show the effectiveness of the method both on model data and on an experimental dataset for which the conformational variability is limited to a defined region within the structure. During our studies of hepatitis C virus (HCV) internal ribosome entry site (IRES) interaction with the human translation initiation factor eIF3, we observed that the IRES RNA included a flexible region holding multiple conformations. While current classification methods were used to produce two-dimensional averages of the complex showing these different conformations, no method existed for relating these averages in three dimensions. Our approach overcame these limitations, giving us structural insight that was previously not possible.

译文

我们描述了一种新颖的方法,用于对具有多个构象状态的结构的类平均值进行排序,以便生成3D重建,以解决样本中存在的构象变异性。该方法假设类平均值之间的相对欧拉角是已知的,然后使用公共线方法从结构的选定视图将任何给定的类与一组不同的构象进行匹配。我们在模型数据和实验数据集上都显示了该方法的有效性,对于这些数据,构象变异性仅限于结构内的定义区域。在我们研究丙型肝炎病毒 (HCV) 内部核糖体进入位点 (IRES) 与人类翻译起始因子eIF3相互作用的过程中,我们观察到IRES RNA包含一个具有多种构象的柔性区域。虽然当前的分类方法用于产生显示这些不同构象的复合物的二维平均值,但不存在将这些平均值在三个维度上关联的方法。我们的方法克服了这些限制,为我们提供了以前无法实现的结构见解。

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