de Forges Hélène, Pilon Antoine, Poüs Christian, Perez Franck
Institut Curie, Paris Cedex 05, France.
Methods Cell Biol. 2013;115:139-53. doi: 10.1016/B978-0-12-407757-7.00010-4.
Microtubules display a very dynamic behavior, and the presence of the guanosine-triphosphate (GTP) cap at the plus ends of microtubules is essential to regulate microtubule dynamics. Dimitrov et al. (2008) showed that GTP-tubulin is present not only at the plus ends but also in discrete locations along the microtubule lattice. These GTP islands were proposed to contribute to rescue events. Studying the localization of GTP-tubulin in microtubules is essential to better comprehend some core aspects in the regulation of microtubule dynamics. In this chapter, we recapitulate essential tools to study the GTP-tubulin using the recombinant antibody MB11 from permeabilized cells to in vitro assays.
微管呈现出非常动态的行为，并且微管正端鸟苷三磷酸（GTP）帽的存在对于调节微管动力学至关重要。迪米特罗夫等人（2008年）表明，GTP微管蛋白不仅存在于正端，还存在于沿微管晶格的离散位置。这些GTP岛被认为有助于救援事件。研究GTP微管蛋白在微管中的定位对于更好地理解微管动力学调节的一些核心方面至关重要。在本章中，我们概述了使用来自通透细胞的重组抗体MB11进行体外测定来研究GTP微管蛋白的基本工具。
