Current Recordings of Ion Channel Proteins Immobilized on Resin Beads
Current Recordings of Ion Channel Proteins Immobilized on Resin Beads
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DOI：
10.1021/ac900286z
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发表时间：
2009-04-15
影响因子：
7.4
通讯作者：
Ide, Toru
中科院分区：
文献类型：
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作者：
Hirano, Minako;Takeuchi, Yuko;Ide, Toru
目前的离子通道电流测量技术是麻烦的，因为它们需要许多步骤和许多时间。当将通道重建成脂质体并将其并入脂质双层时尤其如此。在这里，我们报告了一种新的方法，测量离子通道电流比目前的方法更有效。我们将我们的方法应用于KcsA和MthK通道，通过将它们与具有组氨酸标签的钴亲和凝胶珠结合，然后在珠上形成脂质双层膜。这使得通道纳入双层和通道电流被快速，方便地测量。其效率使得可以用极少量的蛋白质记录电流。此外，通道方向可以由组氨酸标签决定。该方法具有应用于各种通道蛋白和通道研究的潜力。
Current ion channel current measurement techniques are cumbersome, as they require many steps and much time. This is especially true when reconstituting channels into liposomes and incorporating them into lipid bilayers. Here, we report a novel method that measures ion channel current more efficiently than current methods. We applied our method to KcsA and MthK channels by binding them to cobalt affinity gel beads with histidine tags and then forming a lipid bilayer membrane on the bead. This allowed channels to incorporate into the bilayer and channel currents to be measured quickly and easily. The efficiency was such that currents could be recorded with extremely low amounts of protein. In addition, the channel direction could be determined by the histidine tag. This method has the potential to be applied to various channel proteins and channel research in general.