Rectification of voltage-gated plant K+ channels

植物中的电压门控K+通道家族在土壤中钾的吸收及其在植物中的分布起着重要的作用。有趣的是，这个家族的成员在结构上非常相似，但在功能上分为内向整流(Kin)通道和外向整流(Kout)通道，内向整流通道允许植物细胞积累K+，外向整流通道介导K+外流。造成这种分歧的原因尚不清楚。我们产生了一个合成Kin通道(皮肤)和一个合成Kout通道(Skout)，它们有很高的同源性(蛋白质水平上的82.5%，DNA水平上的90%)。人造皮肤和Skout克隆的设计使我们能够进一步利用&gt；30.000.000个不同的皮肤/Skout嵌合体生成一个cDNA文库。该文库将用于识别向内或向外矫正的嵌合体，以进一步增加这两种渠道类型之间的同一性。实验工作伴随着电子分子模拟和动力学模拟。这种结合的方法是新颖的，并将允许对植物K+通道的调节进行详细的结构和功能方面的了解。此外，它将为在计算机上设计具有明确特征的膜蛋白的尝试提供信息数据。

The family of voltage-gated K+ channels in plants plays an important role in the uptake of potassium from the soil and its distribution throughout the plant. Interestingly, members of this family are structurally very similar but functionally they segregate into inward-rectifying (Kin) channels that allow plant cells to accumulate K+ and outward-rectifying (Kout) channels that mediate K+ efflux. The reason for this divergence is unknown. We generated a synthetic Kin channel (SKIN) and a synthetic Kout channel (SKOUT) that share a high rate of identity (82.5% at the protein level, 90% at the DNA level). The design of the synthetic SKIN and SKOUT clones allowed us further to generate a cDNA library with >30.000.000 different SKIN/SKOUT chimeras. This library will be used to identify chimeras that are inward- or outward-rectifying in order to further increase the degree of identity between these two channel types. The experimental work is accompanied by in silico molecular modeling and dynamics simulations. This combined approach is novel and will allow detailed structural and functional insights into the regulation of plant K+ channels. Furthermore, it will provide informative data for the attempt to design membrane proteins with defined features at the computer.