金纳米颗粒和倾斜对来通过冷冻电子显微镜评估蛋白质的柔性。
Gold nanoparticles and tilt pairs to assess protein flexibility by cryo-electron microscopy.
机构信息
Department of Computer Science, Stanford University, Stanford, CA, USA; Department of Electrical Engineering, Stanford University, Stanford, CA, USA.
Department of Computer Science, Stanford University, Stanford, CA, USA.
出版信息
Ultramicroscopy. 2021 Aug;227:113302. doi: 10.1016/j.ultramic.2021.113302. Epub 2021 May 4.
Abstract
A computational method was developed to recover the three-dimensional coordinates of gold nanoparticles specifically attached to a protein complex from tilt-pair images collected by electron microscopy. The program was tested on a simulated dataset and applied to a real dataset comprising tilt-pair images recorded by cryo electron microscopy of RNA polymerase II in a complex with four gold-labeled single-chain antibody fragments. The positions of the gold nanoparticles were determined, and comparison of the coordinates among the tetrameric particles revealed the range of motion within the protein complexes.
摘要
开发了一种计算方法，可从电子显微镜收集的倾斜对图像中恢复特异性附着在蛋白质复合物上的金纳米粒子的三维坐标。该程序在模拟数据集上进行了测试，并应用于包含通过冷冻电子显微镜记录的 RNA 聚合酶 II 与四个金标记的单链抗体片段复合物的倾斜对图像的真实数据集。确定了金纳米粒子的位置，并且四聚体颗粒之间的坐标比较揭示了蛋白质复合物内的运动范围。
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