DNA Origami Scaffolds as Templates for Functional Tetrameric Kir3 K+ Channels
DNA Origami Scaffolds as Templates for Functional Tetrameric Kir3 K+ Channels
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DOI：
10.1002/anie.201709982
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发表时间：
2018-03-01
影响因子：
16.6
通讯作者：
Mori, Yasuo
中科院分区：
文献类型：
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作者：
Kurokawa, Tatsuki;Kiyonaka, Shigeki;Mori, Yasuo
在天然系统中，支架蛋白在将蛋白质组装成复合体以传递信号方面发挥着重要作用。这一概念尚未应用于在人工系统中组装具有功能的跨膜蛋白复合体。为了解决这个问题，DNA折纸有可能作为支架，将蛋白质安排在复合体的特定位置。在这里，我们报道了Kir3K+通道蛋白通过锌指蛋白(ZFP)适配器在DNA折纸支架上的特定位置组装。原子力显微镜和凝胶电泳法证实融合了ZFP的Kir3通道和基于ZFP的接头与DNA折纸发生了特异性结合。此外，带有ZFP结合位点的DNA折纸使异四聚体Kir3通道在HEK293T细胞中诱发的K+通道电流活性增加近两倍。因此，我们的方法提供了一个有用的模板来控制膜蛋白复合体在体外和活细胞中的齐聚状态。
In native systems, scaffolding proteins play important roles in assembling proteins into complexes to transduce signals. This concept is yet to be applied to the assembly of functional transmembrane protein complexes in artificial systems. To address this issue, DNA origami has the potential to serve as scaffolds that arrange proteins at specific positions in complexes. Herein, we report that Kir3 K+ channel proteins are assembled through zinc-finger protein (ZFP)-adaptors at specific locations on DNA origami scaffolds. Specific binding of the ZFP-fused Kir3 channels and ZFP-based adaptors on DNA origami were confirmed by atomic force microscopy and gel electrophoresis. Furthermore, the DNA origami with ZFP binding sites nearly tripled the K+ channel current activity elicited by heterotetrameric Kir3 channels in HEK293T cells. Thus, our method provides a useful template to control the oligomerization states of membrane protein complexes invitro and in living cells.