STRUCTURE

这一倡议的持续目标是确定两个主要的 迁移细胞器、粘连和突起。我们的方法是将各种结构方法结合起来 从相关光学和高分辨电子显微镜到X射线和核磁共振。作为这项努力的一部分，我们正在 开发了一种新的成像概念，使我们能够通过光学显微镜捕捉到 活细胞中的迁移和使用高分辨率电子显微镜显示相同的结构-3D 相关的光学显微镜和电子显微镜(“自上而下的方法”)。个体的高分辨率结构 通过核磁共振、X射线或断层扫描获得蛋白质及其与结合伙伴的详细相互作用 Up Approach“)然后可以通过计算对接到建筑群的三维地图中。在 在联合体的第一阶段，这一范例被应用于突起中肌动蛋白网络结构， 提供了Arp2/3介导的肌动蛋白分支的三维结构和Arp2/3激活的结构基础。我们 还利用我们的新的3D相关的EM和 散斑显微镜，并在整合素存在的情况下确定α-肌动蛋白上的纽蛋白结合部位。统一的 开发了用于分割和分析3D数据的框架，以便能够识别装配 在三维结构中。

The continued goal of this Initiative is to determine the high resolution structures of two major migration organelles, adhesions and protrusions. Our approach is to combine structural methods that range from correlative light and high resolution electron microscopy to X-ray and NMR. As part of this effort, we are developing a new imaging concept that allows us to capture by light microscopy the spatial-temporal aspects of migration in living cells and to visualize the same structures using high resolution electron microscopy - 3D correlative light and electron microscopy ("top down approaches"). High resolution structures of individual proteins and their detailed interactions with binding partners obtained by NMR, X-ray or tomography ("bottom up approaches") can then be computationally docked into the three-dimensional maps of the complexes. In the first phase of the Consortium, this paradigm was applied to the structure of the actin network in protrusions, providing 3D structures of Arp2/3 mediated actin branches and the structural basis for Arp2/3 activation. We also determined the ultrastructure of the lamellipodia of living cells using our new 3D correlative EM and speckle microscopy and determined the vinculin binding site on a-actinin in the presence of integrin. A unified framework for segmentation and analysis of 3-D data was developed to enable the identification of assemblies in 3-D structures.