Unbiased search for structure/function correlates in K+ channels - Exploiting viral molecular evolution to study ion channel functionality

K+通道的功能主要由其结构决定。通常通过比较不同结构的同源物的功能来研究作用于折叠蛋白中的相关复杂和协同结构/功能相关因素。在这里，我们提出了一种新的方法，它应该提供前所未有的高变异性的K+通道同源基因。该策略基于从小球藻病毒丰富的环境样本中分离病毒K+通道并对其进行功能鉴定。几乎所有这些病毒在高选择性压力下都含有一个非常小的K+通道的基因，因为这些通道对宿主感染是必不可少的。在这个项目中，我们将a)从不同的环境样本中分离病毒DNA，b)通过特定的病毒K+通道引物扩增包含的病毒K+通道基因；c)建立丰富的K+通道同源序列文库；d)通过酵母互补在功能上注释这些序列；e)在电子计算机中确定序列中的(共同)进化特征，从而确定进化动力学的边界；f)建立一个除简单结构了解之外的必要分子动力学细节文库，这是发挥作用的K+通道所必需的；以及g)深入了解病毒蛋白质的进化起源。

The function of a K+-channel is mainly determined by its structure. The relevant complex and synergistic structure/function correlates, which operate in the folded protein, are commonly studied by comparing the function of structurally diverse orthologs. Here we propose a new approach, which should provide an unpreceeded high variability of K+-channel orthologs. The strategy is based on isolation and functional characterization of viral K+-channels from environmental samples, where Chlorella viruses are abundant. Nearly all these viruses contain a gene for a very small K+-channel under high selective pressure, as these channels are essential for host infection. In this project we will a) isolate viral DNA from diverse environmental samples, b) amplify contained viral K+-channel genes by specific viral K+-channel primers; c) build a rich sequence library of K+-channel orthologs; d) annotated these functionally by yeast complementation; e) determine in silico the (co)evolutionary signatures in the sequences and thus boundaries of evolutionary dynamics; f) build a library of necessary molecular dynamical details beyond simple structural insight, which are necessary for functional K+-channels; and g) gain insight into the evolutionary origins of viral proteins.