(-)-Epigallocatechin Gallate is a Noncompetitive Inhibitor of NAD Kinase.

ABSTRACT: 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.

SUBMITTER: Liu T 

PROVIDER: S-EPMC9661698 | biostudies-literature | 2022 Nov

REPOSITORIES: biostudies-literature