Elie-Caille Céline, Severin Fedor, Helenius Jonne, Howard Jonathon, Muller Daniel J, Hyman A A
Department of Cellular Machines, BioTechnological Center, University of Technology Dresden, Tatzberg 49, D-01307 Dresden, Germany.
Curr Biol. 2007 Oct 23;17(20):1765-70. doi: 10.1016/j.cub.2007.08.063. Epub 2007 Oct 4.
Microtubules exist in dynamic equilibrium, growing and shrinking by the addition or loss of tubulin dimers from the ends of protofilaments. The hydrolysis of GTP in beta-tubulin destabilizes the microtubule lattice by increasing the curvature of protofilaments in the microtubule and putting strain on the lattice. The observation that protofilament curvature depends on GTP hydrolysis suggests that microtubule destabilizers and stabilizers work by modulating the curvature of the microtubule lattice itself. Indeed, the microtubule destabilizer MCAK has been shown to increase the curvature of protofilaments during depolymerization. Here, we show that the atomic force microscopy (AFM) of individual tubulin protofilaments provides sufficient resolution to allow the imaging of single protofilaments in their native environment. By using this assay, we confirm previous results for the effects of GTP hydrolysis and MCAK on the conformation of protofilaments. We go on to show that taxol stabilizes microtubules by straightening the GDP protofilament and slowing down the transition of protofilaments from straight to a curved configuration.
微管处于动态平衡中,通过原纤维末端微管蛋白二聚体的添加或丢失而生长和收缩。β-微管蛋白中GTP的水解通过增加微管中原纤维的曲率并对晶格施加应变来使微管晶格不稳定。原纤维曲率取决于GTP水解这一观察结果表明,微管去稳定剂和稳定剂通过调节微管晶格本身的曲率起作用。事实上,微管去稳定剂MCAK已被证明在解聚过程中会增加原纤维的曲率。在这里,我们表明单个微管蛋白原纤维的原子力显微镜(AFM)提供了足够的分辨率,以允许在其天然环境中对单个原纤维进行成像。通过使用该测定法,我们证实了先前关于GTP水解和MCAK对原纤维构象影响的结果。我们接着表明,紫杉醇通过拉直GDP原纤维并减缓原纤维从直构象到弯曲构象的转变来稳定微管。
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