Interenzyme substrate diffusion for an enzyme cascade organized on spatially addressable DNA nanostructures.
Interenzyme substrate diffusion for an enzyme cascade organized on spatially addressable DNA nanostructures.
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DOI：
10.1021/ja300897h
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发表时间：
2012-03-28
影响因子：
15
通讯作者：
Yan, Hao
中科院分区：
文献类型：
--
作者：
Fu, Jinglin;Liu, Minghui;Liu, Yan;Woodbury, Neal W.;Yan, Hao
空间可寻址的DNA纳米结构促进了具有精确控制模式的异质元件的自组装。在这里，我们在特定的DNA折纸瓷砖上组织了离散的Gox/HRP酶对，并控制了酶间的间距和位置。酶之间的距离从10 nm到65 nm系统变化，并对相应的活性进行了评估。这项研究揭示了与组装的酶对相关的两个不同的距离依赖的动力学过程。观察到那些酶排列紧密的组装体的活性显著增强，而相距仅20 nm的酶的活性显著下降。进一步增加间距会导致较弱的距离相关性。结合扩散模型，结果表明，中间体在溶液中的布朗扩散控制了较远的酶对活性的变化，而中间体在连接的蛋白质表面上的尺寸受限扩散有助于提高紧密间距的Gox/HRP组装的活性。为了进一步测试沿蛋白质表面的有限维扩散的作用，在Gox和HRP之间插入了一座非催化的蛋白质桥来连接它们的水合壳。这大大提高了酶对的活性。
Spatially addressable DNA nanostructures facilitate the self-assembly of heterogeneous elements with precisely controlled patterns. Here we organized discrete GOx/HRP enzyme pairs on specific DNA origami tiles with controlled inter-enzyme spacing and position. The distance between enzymes was systematically varied from 10 nm to 65 nm and the corresponding activities were evaluated. The study revealed two different distance dependent kinetic processes associated with the assembled enzyme pairs. Strongly enhanced activity was observed for those assemblies in which the enzymes were closely spaced, while the activity dropped dramatically for enzymes as little as 20 nm apart. Increasing the spacing further resulted in a much weaker distance dependence. Combined with diffusion modeling, the results suggest that Brownian diffusion of intermediates in solution governed the variations in activity for more distant enzyme pairs, while dimensionally-limited diffusion of intermediates across connected protein surfaces contributed to the enhancement in activity for closely spaced GOx/HRP assemblies. To further test the role of limited dimensional diffusion along protein surfaces, a noncatalytic protein bridge was inserted between GOx and HRP to connect their hydration shells. This resulted in substantially enhanced activity of the enzyme pair.
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影响因子：
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通讯作者：
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