Autonomous loading, transport, and unloading of specified cargoes by using DNA hybridization and biological motor-based motility
Autonomous loading, transport, and unloading of specified cargoes by using DNA hybridization and biological motor-based motility
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DOI：
10.1002/smll.200700528
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发表时间：
2008-04-01
期刊：
影响因子：
13.3
通讯作者：
Sutoh, Kazuo
中科院分区：
文献类型：
--
作者：
Hiyama, Satoshi;Inoue, Takeshi;Sutoh, Kazuo
生物马达（如运动蛋白）在没有外界刺激的情况下装载/卸载特定类型的货物（如囊泡），并利用真核细胞内（体内）三磷酸腺苷（ATP）水解的能量，沿着细胞骨架细丝（如微管）运输这些货物由于这些惊人的生物能力，即自主装载/卸载和特定货物的运输，在纳米或细胞级系统和应用中，将驱动蛋白和mt结合到人工制造的（体外）转运体和致动器中有相当大的兴趣。[2-19]我们的目标是创建一个重组的分子运输系统，该系统使用DNA杂交装载/卸载指定货物，并使用运动蛋白上MT运动的反向几何形状运输它们（体外滑行试验）。我们的研究结果表明，特定的货物在没有外部刺激的情况下被装载/卸载到滑行mt上并由mt运输。该通讯首次展示了在体外自主装卸和运输特定货物。自从肌动蛋白上的肌动蛋白运动的反向几何结构（体外滑动实验）被成功证明以来，重建的肌动蛋白运动受到越来越多的关注为了将MT运动作为一种分子运输系统，需要在给定的装载位置将指定的货物装载到滑动MT上，将装载的MT定向运输到给定的卸载位置，然后在没有外部刺激的情况下将货物从卸载位置的MT上卸下。这种特定货物的自主装卸和定向运输可能有助于创建高度小型化的芯片上系统，如分子分拣器、分子传感器和分子通信系统。[16,17]为了在分子运输系统中实现货物的定向运输，现有的技术使用预先配置的微光刻轨道[2-5]可能适用于控制装载到滑动MTs上的货物的运输方向。然而，对于货物的自主装卸，需要一种新的方法，因为现有的系统利用亲和素-生物素[3,9]或抗原-抗体[10]，[11]由于绑定紧密，绑定可能不适合货物卸载。其他现有的系统需要外部刺激，如紫外线照射、[13,14]配体补充、[15]限制性内切酶消化或温度波动[18]，才能从滑动的mt中卸载货物；因此，这些系统是非自治的。
Biological motors (eg, kinesins) load/unload particular types of cargoes (eg, vesicles) without external stimuli, and transport them along cytoskeletal filaments such as microtubules (MTs) by using the energy of adenosine triphosphate (ATP) hydrolysis within eukaryotic cells (in vivo).[1] Because of these amazing biological capabilities, that is, autonomous loading/unloading and transport of specified cargoes, there is considerable interest in incorporating kinesins and MTs into artificially created (in vitro) transporters and actuators in nanometer-or cell-scale systems and applications.[2–19] We aimed to create a reconstituted molecular transport system that loads/unloads specified cargoes using DNA hybridization and transports them using the reverse geometry of MT motility on kinesins (in vitro gliding assay). Our results show that specified cargoes were loaded/unloaded onto/from gliding MTs and transported by the MTs without external stimuli. This Communication is the first to demonstrate autonomous loading/unloading and transport of specified cargoes in vitro.Reconstituted MT motility has received increasing attention ever since a reverse geometry of MT motility on kinesins (in vitro gliding assay) was successfully demonstrated.[20] To use MT motility as a molecular transport system, it is necessary to load specified cargoes onto gliding MTs at a given loading site, directionally transport the loaded MTs towards a given unloading site, and then unload the cargoes from the MTs at the unloading site without external stimuli. Such autonomous loading/unloading and directional transport of specified cargoes may help create highly miniaturized on-chip systems such as molecular sorters, molecular sensors, and molecular communication systems.[16, 17] For achieving the directional transport of cargoes in a molecular transport system, existing techniques using preconfigured microlithographic tracks [2–5] may be applicable for controlling the transport direction of cargoes loaded onto gliding MTs. However, for autonomous loading/unloading of cargoes a new approach is required, because existing systems that utilize avidin–biotin [3, 9] or antigen–antibody [10, 11] bindings may not be suitable for cargo unloading owing to their tight bindings. Other existing systems require external stimuli such as UV exposure,[13, 14] ligand supplementation,[15] restriction enzyme digestion, or temperature ffuctuation [18] to unload cargoes from gliding MTs; thus making those systems non-autonomous.