Structural biology in situ - the potential of subtomogram averaging
Structural biology in situ - the potential of subtomogram averaging
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DOI：
10.1016/j.sbi.2013.02.003
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发表时间：
2013-04-01
影响因子：
6.8
通讯作者：
Briggs, John A. G.
中科院分区：
文献类型：
--
作者：
Briggs, John A. G.
冷冻电子断层扫描提供细胞、细胞器或病毒的低分辨率3D视图。存在于多个副本中的大分子复合物可以随后在3D重建（断层图像）内识别，计算提取，并平均以获得更高分辨率的3D结构，以及它们的空间分布图。这种方法，称为subtomogram平均或子体积平均，允许大分子复合物的结构，以解决在原位。最近的应用程序提供了原位结构数据的分辨率为2-4纳米的样品，包括多聚核糖体，核孔，囊泡外套，和病毒表面蛋白。在这里，我描述的方法和讨论的局限性，进步和最近的应用。我推测该方法将如何以更高的分辨率解决更多的结构，从而实现原位结构生物学。
Cryo-electron tomography provides low-resolution 3D views of cells, organelles, or viruses. Macromolecular complexes present in multiple copies can be subsequently identified within the 3D reconstruction (the tomogram), computationally extracted, and averaged to obtain higher resolution 3D structures, as well as a map of their spatial distribution. This method, called subtomogram averaging or subvolume averaging, allows structures of macromolecular complexes to be resolved in situ. Recent applications have provided in situ structural data at resolutions of 2-4 nm on samples including polysomes, nuclear pores, vesicle coats, and viral surface proteins. Here I describe the method and discuss limitations, advances and recent applications. I speculate how the method will solve more structures at higher resolution, allowing in situ structural biology.