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Strain through the neck linker ensures processive runs: a DNA-kinesin hybrid nanomachine study

机译:通过颈部连接器的应变确保了连续运行:DNA-驱动蛋白混合纳米机研究

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摘要

The motor protein kinesin has two heads and walks along microtubules processively using energy derived from ATP. However, how kinesin heads are coordinated to generate processive movement remains elusive. Here we created a hybrid nanomachine (DNA-kinesin) using DNA as the skeletal structure and kinesin as the functional module. Single molecule imaging of DNA-kinesin hybrid allowed us to evaluate the effects of both connect position of the heads (N, C-terminal or Mid position) and sub-nanometer changes in the distance between the two heads on motility. Our results show that although the native structure of kinesin is not essential for processive movement, it is the most efficient. Furthermore, forward bias by the power stroke of the neck linker, a 13-amino-acid chain positioned at the C-terminus of the head, and internal strain applied to the rear of the head through the neck linker are crucial for the processive movement. Results also show that the internal strain coordinates both heads to prevent simultaneous detachment from the microtubules. Thus, the inter-head coordination through the neck linker facilitates long-distance walking.
机译:运动蛋白驱动蛋白具有两个头部,并利用源自ATP的能量依次沿着微管运动。然而,如何协调驱动蛋白头以产生进行性运动仍然是未知的。在这里,我们创建了一个混合的纳米机器(DNA驱动蛋白),使用DNA作为骨架结构,并以驱动蛋白作为功能模块。 DNA-驱动蛋白杂合体的单分子成像使我们能够评估头部的连接位置(N,C端或中间位置)以及两个头部之间距离的亚纳米变化对运动性的影响。我们的结果表明,尽管驱动蛋白的天然结构对于进行性运动不是必需的,但它是最有效的。此外,颈部连接器的动力冲程产生的前向偏置,位于头部C端的13个氨基酸链以及通过颈部连接器施加到头部后部的内部应变对于进行性运动至关重要。结果还表明,内部应变协调了两个头部,以防止同时从微管上脱落。因此,通过颈部连接器的头部间协调有助于长距离行走。

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