The force required to remove tubulin from the microtubule lattice by pulling on its α-tubulin C-terminal tail.
The force required to remove tubulin from the microtubule lattice by pulling on its α-tubulin C-terminal tail.
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DOI：
10.1038/s41467-022-31069-x
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发表时间：
2022-06-25
影响因子：
16.6
通讯作者：
中科院分区：
文献类型：
--
作者：
切断的酶和分子马达通过对埋藏在晶格中的亚单位施加机械力，从微管的壁上提取微管蛋白。然而，需要多大的力量才能从微管中移除微管蛋白，也不知道亚基被移除的途径。利用一种特定部位的功能化方法，我们用光钳对α-微管蛋白的C-末端施加作用力，发现从微管管壁中提取微管蛋白需要大约30pN的力。此外，我们发现部分去折叠是微管蛋白去除的中间步骤。去折叠力和提取力类似于AAA-去折叠酶产生的力。最后，我们证明了三种Kinesin-1马达蛋白也可以从微管晶格中提取微管蛋白。我们的结果首次对微管蛋白如何响应施加在其α-微管蛋白C末端的机械力进行了实验研究，并对切断酶的机制和微管的稳定性具有重要意义。微管的构建块微管蛋白可以通过机械力从微管壁上移除。使用单分子方法，作者证明了微管蛋白在去除之前部分展开，并确定了微管蛋白的抽取力。
Severing enzymes and molecular motors extract tubulin from the walls of microtubules by exerting mechanical force on subunits buried in the lattice. However, how much force is needed to remove tubulin from microtubules is not known, nor is the pathway by which subunits are removed. Using a site-specific functionalization method, we applied forces to the C-terminus of α-tubulin with an optical tweezer and found that a force of ~30 pN is required to extract tubulin from the microtubule wall. Additionally, we discovered that partial unfolding is an intermediate step in tubulin removal. The unfolding and extraction forces are similar to those generated by AAA-unfoldases. Lastly, we show that three kinesin-1 motor proteins can also extract tubulin from the microtubule lattice. Our results provide the first experimental investigation of how tubulin responds to mechanical forces exerted on its α-tubulin C-terminal tail and have implications for the mechanisms of severing enzymes and microtubule stability. Tubulin, the building blocks of microtubules, can be removed from the microtubule wall by mechanical forces. Using single-molecule methods, the authors show that tubulin partially unfolds prior to its removal and determined the tubulin-extraction force.
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