Department of Synthetic Chemistry and Biological Chemistry, Graduate School of Engineering, Kyoto University, Katsura, Nishikyo-ku, Kyoto 615-8510, Japan.
Department of Technology and Ecology, Hall of Global Environmental Studies, Kyoto University, Katsura, Nishikyo-ku, Kyoto 615-8510, Japan.
Nat Chem. 2016 Oct;8(10):958-67. doi: 10.1038/nchem.2554. Epub 2016 Jun 27.
The controlled activation of proteins in living cells is an important goal in protein-design research, but to introduce an artificial activation switch into membrane proteins through rational design is a significant challenge because of the structural and functional complexity of such proteins. Here we report the allosteric activation of two types of membrane-bound neurotransmitter receptors, the ion-channel type and the G-protein-coupled glutamate receptors, using coordination chemistry in living cells. The high programmability of coordination chemistry enabled two His mutations, which act as an artificial allosteric site, to be semirationally incorporated in the vicinity of the ligand-binding pockets. Binding of Pd(2,2'-bipyridine) at the allosteric site enabled the active conformations of the glutamate receptors to be stabilized. Using this approach, we were able to activate selectively a mutant glutamate receptor in live neurons, which initiated a subsequent signal-transduction pathway.
在活细胞中控制蛋白质的激活是蛋白质设计研究的一个重要目标，但由于这些蛋白质的结构和功能复杂性，通过合理设计将人工激活开关引入膜蛋白是一项重大挑战。在这里，我们报告了使用活细胞中的配位化学对两种类型的膜结合神经递质受体（离子通道型和 G 蛋白偶联谷氨酸受体）进行别构激活。配位化学的高度可编程性使两个 His 突变（作为人工别构位点）半理性地整合到配体结合口袋附近。在别构位点结合 Pd(2,2'-联吡啶)可稳定谷氨酸受体的活性构象。通过这种方法，我们能够选择性地激活活神经元中的突变型谷氨酸受体，从而启动后续的信号转导途径。
