(-)-Epigallocatechin Gallate is a Noncompetitive Inhibitor of NAD Kinase.
Tonghai LiuWenjia ShiYiluan DingQiqi WuBei ZhangNaixia ZhangMingliang WangDaohai DuHao ZhangBo HanDe-An GuoJie ZhengQi LiCheng LuoPublished in: ACS medicinal chemistry letters (2022)
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.
Keyphrases
- wild type
- mass spectrometry
- high throughput
- multiple sclerosis
- reactive oxygen species
- liquid chromatography
- ms ms
- small molecule
- cell death
- dna damage
- signaling pathway
- genome wide
- gene expression
- high performance liquid chromatography
- cancer therapy
- tyrosine kinase
- binding protein
- copy number
- dna methylation
- dna binding
- capillary electrophoresis
- gas chromatography
- data analysis
- solid phase extraction
- fluorescent probe