Defining and Exploiting Hypersensitivity Hotspots to Facilitate Abscisic Acid Agonist Optimization
Defining and Exploiting Hypersensitivity Hotspots to Facilitate Abscisic Acid Agonist Optimization
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DOI：
10.1021/acschembio.8b00955
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发表时间：
2019-03-01
影响因子：
4
通讯作者：
Cutler, Sean R.
中科院分区：
文献类型：
--
作者：
Elzinga, Dezi;Sternburg, Erin;Cutler, Sean R.
吡拉菌素抗性1（PYR1）及其相关的脱落酸（阿坝）受体是调控植物耐旱性的新靶点。在这里，我们确定并使用PYR1超敏突变体来定义配体结合热点，并表明这些可以指导激动剂效力的改善。定义的一个热点残基A160是被阿坝的C6甲基或合成激动剂喹那菌素（QB）的甲苯甲酰基甲基占据的口袋的一部分。合成了一系列在甲苯甲酰基位置取代的QB类似物，并在体外提供高达10倍的活性增益。此外，我们证明，超敏受体可用于提高灵敏度的哺乳动物细胞ABA调节的转录电路的三个数量级。总的来说，我们的数据表明，在配体结合口袋中的超敏反应位点的系统映射可以帮助指导配体优化和调整工程受体的敏感性。
Pyrabactin resistance 1 (PYR1) and related abscisic acid (ABA) receptors are new targets for manipulating plant drought tolerance. Here, we identify and use PYR1 hypersensitive mutants to define ligand binding hotspots and show that these can guide improvements in agonist potency. One hotspot residue defined, A160, is part of a pocket that is occupied by ABA's C6 methyl or by the toluyl methyl of the synthetic agonist quinabactin (QB). A series of QB analogues substituted at the toluyl position were synthesized and provide up to 10-fold gain in activity in vitro. Furthermore, we demonstrate that hypersensitive receptors can be used to improve the sensitivity of a previously described mammalian cell ABA-regulated transcriptional circuit by three orders of magnitude. Collectively, our data show that the systematic mapping of hypersensitivity sites in a ligand-binding pocket can help guide ligand optimization and tune the sensitivity of engineered receptors.