(-)-Epigallocatechin Gallate is a Noncompetitive Inhibitor of NAD Kinase.
(-)-Epigallocatechin Gallate is a Noncompetitive Inhibitor of NAD Kinase.
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DOI：
10.1021/acsmedchemlett.2c00163
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发表时间：
2022-10
影响因子：
4.2
通讯作者：
Tonghai Liu;Wenjia Shi;Yi-luan Ding;Qiqi Wu;Bei Zhang;Naixia Zhang;Mingliang Wang;Daohai Du;Hao Zhang;Bo Han;D. Guo;Jie Zheng;Qi Li;C. Luo
中科院分区：
文献类型：
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作者：
Tonghai Liu;Wenjia Shi;Yi-luan Ding;Qiqi Wu;Bei Zhang;Naixia Zhang;Mingliang Wang;Daohai Du;Hao Zhang;Bo Han;D. Guo;Jie Zheng;Qi Li;C. Luo
烟酰胺腺嘌呤二核苷酸激酶（Nicotinamide adenine dinucleotide kinase， NADK）控制细胞内NADPH的含量，为大分子和抗ros的合成提供还原能力。此外，NADK被认为是KRAS突变的合成致死基因。为了发现nadk靶向探针，建立了高通量筛选实验，并对其Z因子为0.71进行了优化。天然产物(-)-表没食子儿茶素没食子酸酯（EGCG）是NADK的非竞争性抑制剂，其K i = 3.28±0.32 μΜ。EGCG与NADK的直接结合是通过几种生物物理方法确定的，包括核磁共振光谱、表面等离子体共振（SPR）测定和氢-氘交换质谱（HDX-MS）。SPR测定的K d为1.78±1.15 μΜ。HDX-MS实验表明，EGCG结合在NADK的非底物结合位点。结合模式预测和衍生物活性分析揭示了EGCG和NADK之间潜在的构效关系。此外，EGCG可以特异性抑制KRAS突变型肺癌细胞株的增殖，而不影响KRAS野生型肺癌细胞株。
Nicotinamide adenine dinucleotide kinase (NADK) controls the intracellular NADPH content and provides reducing power for the synthesis of macromolecules and anti-ROS. Moreover, NADK is considered to be a synthetic lethal gene for KRAS mutations. To discover NADK-targeted probes, a high-throughput screening assay was established and optimized with a Z factor of 0.71. The natural product (-)-epigallocatechin gallate (EGCG) was found to be a noncompetitive inhibitor of NADK with K i = 3.28 ± 0.32 μΜ. The direct binding of EGCG to NADK was determined by several biophysical methods, including NMR spectroscopy, surface plasmon resonance (SPR) assay, and hydrogen-deuterium exchange mass spectrometry (HDX-MS). The SPR assay showed a K d of 1.78 ± 1.15 μΜ. The HDX-MS experiment showed that EGCG was bound at the non-substrate-binding sites of NADK. Besides, binding mode prediction and derivative activity analysis revealed a potential structure-activity relationship between EGCG and NADK. Furthermore, EGCG can specifically inhibit the proliferation of KRAS-mutated lung cancer cell lines without affecting KRAS wild-type lung cancer cell lines.