Ku Siou, Messaoudi Cédric, Guyomar Charlotte, Kervrann Charles, Chrétien Denis
Univ Rennes, CNRS, IGDR (Institut de genetique et developpement de Rennes) - UMR 6290, F-35000 Rennes, France.
Institut Curie, PSL Research University, CNRS UMS 2016, F-91401 Orsay, France.
Bio Protoc. 2020 Nov 5;10(21):e3814. doi: 10.21769/BioProtoc.3814.
The α-β tubulin heterodimer undergoes subtle conformational changes during microtubule assembly. These can be modulated by external factors, whose effects on microtubule structure can be characterized on 2D views obtained by cryo-electron microscopy. Analysis of microtubule images is facilitated if they are straight enough to interpret and filter their image Fourier transform, which provide useful information concerning the arrangement of tubulin molecules inside the microtubule lattice. Here, we describe the use of the TubuleJ software to straighten microtubules and determine their lattice parameters. Basic 3D reconstructions can be performed to evaluate the relevance of these parameters. This approach can be used to analyze the effects of nucleotide analogues, drugs or MAPs on microtubule structure, or to select microtubule images prior to high-resolution 3D reconstructions.
α-β微管蛋白异二聚体在微管组装过程中会发生细微的构象变化。这些变化可由外部因素调节,其对微管结构的影响可通过冷冻电子显微镜获得的二维视图进行表征。如果微管图像足够笔直以便解释并对其图像傅里叶变换进行滤波,那么微管图像分析会更容易,这能提供有关微管晶格内微管蛋白分子排列的有用信息。在此,我们描述了使用TubuleJ软件来拉直微管并确定其晶格参数。可以进行基本的三维重建以评估这些参数的相关性。这种方法可用于分析核苷酸类似物、药物或微管相关蛋白对微管结构的影响,或在高分辨率三维重建之前选择微管图像。