Pterostilbene-mediated Nrf2 activation: Mechanistic insights on Keap1:Nrf2 interface
Pterostilbene-mediated Nrf2 activation: Mechanistic insights on Keap1:Nrf2 interface
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DOI：
10.1016/j.bmc.2016.05.011
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发表时间：
2016-08-15
影响因子：
3.5
通讯作者：
Ramkumar, Kunka Mohanram
中科院分区：
文献类型：
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作者：
Bhakkiyalakshmi, Elango;Dineshkumar, Kesavan;Ramkumar, Kunka Mohanram
Keap 1-Nrf 2蛋白质相互作用（PPI）抑制剂的发现为开发新型抗应激先导分子提供了一个很有前途的策略。因此，Keap 1-Nrf 2系统在氧化/亲电应激相关疾病中起重要作用。我们早期的研究确定了紫檀芪（PTS），白藜芦醇的天然类似物，作为一种有效的Nrf 2激活剂和Keap 1-Nrf 2 PPI抑制剂，通过荧光素酶互补试验进行评估。本研究通过核转位实验和ARE荧光素酶报告基因实验进一步验证了PTS在Nrf 2激活和ARE驱动下游靶基因表达中的作用。此外，荧光素酶互补测定确定PTS以剂量和时间依赖性方式抑制Keap 1-Nrf 2 PPI。通过分子对接和动力学模拟的计算研究表明，PTS直接与Keap 1的kelch结构域的碱性氨基酸相互作用，并干扰了Keap 1-Nrf 2的相互作用模式。该手稿不仅显示了Keap 1-Nrf 2蛋白的结合决定簇，而且还提供了PTS的Nrf 2激活潜力的机制见解。(C)2016爱思唯尔有限公司版权所有
The discovery of Keap1-Nrf2 protein-protein interaction (PPI) inhibitors has become a promising strategy to develop novel lead molecules against variety of stress. Hence, Keap1-Nrf2 system plays an important role in oxidative/electrophilic stress associated disorders. Our earlier studies identified pterostilbene (PTS), a natural analogue of resveratrol, as a potent Nrf2 activator and Keap1-Nrf2 PPI inhibitor as assessed by luciferase complementation assay. In this study, we further identified the potential of PTS in Nrf2 activation and ARE-driven downstream target genes expression by nuclear translocation experiments and ARE-luciferase reporter assay, respectively. Further, the luciferase complementation assay identified that PTS inhibits Keap1-Nrf2 PPI in both dose and time-dependent manner. Computational studies using molecular docking and dynamic simulation revealed that PTS directly interacts with the basic amino acids of kelch domain of Keap1 and perturb Keap1-Nrf2 interaction pattern. This manuscript not only shows the binding determinants of Keap1-Nrf2 proteins but also provides mechanistic insights on Nrf2 activation potential of PTS. (C) 2016 Elsevier Ltd. All rights reserved.