Conformational changes in tubulin in GMPCPP and GDP-taxol microtubules observed by cryoelectron microscopy.
Conformational changes in tubulin in GMPCPP and GDP-taxol microtubules observed by cryoelectron microscopy.
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DOI：
10.1083/jcb.201201161
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发表时间：
2012-08-06
期刊：
影响因子：
--
通讯作者：
Hirokawa N
中科院分区：
文献类型：
--
作者：
Yajima H;Ogura T;Nitta R;Okada Y;Sato C;Hirokawa N
冷冻电子显微镜揭示了GMPCPP和GDP-紫杉醇微管之间的微管蛋白之间的接触的构象变化。微管是动态聚合物，在生长和收缩阶段之间随机切换。微管动力学受β-微管蛋白水解三磷酸鸟苷（GTP）的调节，但这种调节的机制仍然难以捉摸，因为高分辨率的微管结构只显示了鸟苷二磷酸（GDP）状态。本文提出了一种新的冷冻电镜（cryo-EM）图像重建算法，在8.8 μ m分辨率下，对GTP类似物5′-α，β-亚甲基二膦酸鸟苷酯（GMPCPP）稳定的微管进行了冷冻电镜（cryo-EM）图像重建。与GTP结合的微管蛋白相关物如γ-微管蛋白和细菌微管蛋白的晶体结构相比，在微管蛋白之间的接触处（沿着原丝和相邻原丝之间），GMPCPP和GDP-紫杉醇微管之间检测到显著的变化，这有助于微管的稳定性。这些发现是一致的结构可塑性或晶格模型，并建议的结构基础，不仅为微管动力学的调节机制，但也为识别的核苷酸状态的微管由几个微管结合蛋白，如EB 1或驱动蛋白。
Cryoelectron microscopy reveals conformational changes at the contacts between tubulins between GMPCPP and GDP-taxol microtubules. Microtubules are dynamic polymers that stochastically switch between growing and shrinking phases. Microtubule dynamics are regulated by guanosine triphosphate (GTP) hydrolysis by β-tubulin, but the mechanism of this regulation remains elusive because high-resolution microtubule structures have only been revealed for the guanosine diphosphate (GDP) state. In this paper, we solved the cryoelectron microscopy (cryo-EM) structure of microtubule stabilized with a GTP analogue, guanylyl 5′-α,β-methylenediphosphonate (GMPCPP), at 8.8-Å resolution by developing a novel cryo-EM image reconstruction algorithm. In contrast to the crystal structures of GTP-bound tubulin relatives such as γ-tubulin and bacterial tubulins, significant changes were detected between GMPCPP and GDP-taxol microtubules at the contacts between tubulins both along the protofilament and between neighboring protofilaments, contributing to the stability of the microtubule. These findings are consistent with the structural plasticity or lattice model and suggest the structural basis not only for the regulatory mechanism of microtubule dynamics but also for the recognition of the nucleotide state of the microtubule by several microtubule-binding proteins, such as EB1 or kinesin.
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影响因子：
3.3
作者：
HYMAN, AA;SALSER, S;MITCHISON, TJ
通讯作者：
MITCHISON, TJ
DOI：
10.1083/jcb.201104034
发表时间：
2011-07-25
期刊：
The Journal of cell biology
影响因子：
--
作者：
Nakata T;Niwa S;Okada Y;Perez F;Hirokawa N
通讯作者：
Hirokawa N
DOI：
10.1073/pnas.1014758108
发表时间：
2011-03-08
影响因子：
11.1
作者：
Maurer, Sebastian P.;Bieling, Peter;Surrey, Thomas
通讯作者：
Surrey, Thomas
影响因子：
5.6
作者：
Bodey, Andrew J.;Kikkawa, Masahide;Moores, Carolyn A.
通讯作者：
Moores, Carolyn A.
影响因子：
11.4
作者：
Kikkawa, Masahide;Hirokawa, Nobutaka
通讯作者：
Hirokawa, Nobutaka