Department of Biochemistry, University of Geneva, 1211 Geneva, Switzerland.
National Center for Competence in Research Chemical Biology, University of Geneva, 1211 Geneva, Switzerland.
STAR Protoc. 2022 Apr 18;3(2):101320. doi: 10.1016/j.xpro.2022.101320. eCollection 2022 Jun 17.
Microtubules are dynamic polymers where tubulin exchanges not only at the ends but also all along the microtubule shaft. reconstitutions are a vital approach to study microtubule tip dynamics, while direct observation of shaft dynamics is challenging. Here, we describe a dual-color assay to visualize microtubule shaft dynamics using purified, labeled bovine brain tubulin. With this assay, we can quantitatively address how proteins or small molecules impact the dynamics at the microtubule shaft. For complete details on the use and execution of this protocol, please refer to Andreu-Carbó et al. (2022).
微管是动态聚合物，其中微管蛋白不仅在末端交换，而且沿着整个微管轴交换。重构是研究微管尖端动力学的重要方法，而直接观察轴动力学具有挑战性。在这里，我们描述了一种使用纯化的、标记的牛脑微管蛋白可视化微管轴动力学的双色测定法。使用该测定法，我们可以定量地研究蛋白质或小分子如何影响微管轴的动力学。有关该方案使用和执行的完整详细信息，请参阅 Andreu-Carbó 等人（2022 年）。
