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直接测量从正在解聚的微管末端向外卷曲的原丝中的构象应变能。

Direct measurement of conformational strain energy in protofilaments curling outward from disassembling microtubule tips.

作者信息

Driver Jonathan W, Geyer Elisabeth A, Bailey Megan E, Rice Luke M, Asbury Charles L

机构信息

Department of Physiology and Biophysics, University of Washington, Seattle, United States.

Department of Biophysics, UT Southwestern Medical Center, Dallas, United States.

出版信息

Elife. 2017 Jun 19;6:e28433. doi: 10.7554/eLife.28433.

Abstract

Disassembling microtubules can generate movement independently of motor enzymes, especially at kinetochores where they drive chromosome motility. A popular explanation is the 'conformational wave' model, in which protofilaments pull on the kinetochore as they curl outward from a disassembling tip. But whether protofilaments can work efficiently via this spring-like mechanism has been unclear. By modifying a previous assay to use recombinant tubulin and feedback-controlled laser trapping, we directly demonstrate the spring-like elasticity of curling protofilaments. Measuring their mechanical work output suggests they carry ~25% of the energy of GTP hydrolysis as bending strain, enabling them to drive movement with efficiency similar to conventional motors. Surprisingly, a β-tubulin mutant that dramatically slows disassembly has no effect on work output, indicating an uncoupling of disassembly speed from protofilament strain. These results show the wave mechanism can make a major contribution to kinetochore motility and establish a direct approach for measuring tubulin mechano-chemistry.

摘要

微管的拆解能够独立于运动酶产生运动,尤其是在驱动染色体运动的动粒处。一种流行的解释是“构象波”模型,在该模型中,原纤维从拆解末端向外卷曲时会拉动动粒。但原纤维是否能通过这种类似弹簧的机制高效工作尚不清楚。通过改进先前的实验,使用重组微管蛋白和反馈控制的激光捕获技术,我们直接证明了卷曲原纤维的弹簧状弹性。测量它们的机械功输出表明,它们以弯曲应变的形式携带了约25%的GTP水解能量,使其能够以与传统马达相似的效率驱动运动。令人惊讶的是,一种显著减缓拆解速度的β-微管蛋白突变体对功输出没有影响,这表明拆解速度与原纤维应变之间没有关联。这些结果表明,波动机制可以对动粒运动做出重大贡献,并建立了一种测量微管蛋白机械化学的直接方法。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/973c/5515574/78be58337c35/elife-28433-fig1.jpg

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