Allosteric activation of membrane-bound glutamate receptors using coordination chemistry within living cells
Allosteric activation of membrane-bound glutamate receptors using coordination chemistry within living cells
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DOI：
10.1038/nchem.2554
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发表时间：
2016-10-01
期刊：
影响因子：
21.8
通讯作者：
Hamachi, Itaru
中科院分区：
文献类型：
--
作者：
Kiyonaka, Shigeki;Kubota, Ryou;Hamachi, Itaru
活细胞中蛋白质的受控激活是蛋白质设计研究的重要目标，但由于这些蛋白质的结构和功能的复杂性，通过合理的设计将人工激活开关引入膜蛋白是一个重大的挑战。在这里，我们报告的变构激活两种类型的膜结合的神经递质受体，离子通道型和G-蛋白偶联谷氨酸受体，在活细胞中使用配位化学。配位化学的高度可编程性使两个His突变（其充当人工变构位点）能够半理性地并入配体结合口袋附近。Pd（2，2 '-联吡啶）在变构位点的结合使谷氨酸受体的活性构象得以稳定。使用这种方法，我们能够选择性地激活活神经元中的突变型谷氨酸受体，从而启动随后的信号转导途径。
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.