(-)-Epigallocatechin-3-gallate, a potential inhibitor to human dicarbonyl/L-xylulose reductase

• Published in: Journal of biochemistry (Tokyo) Vol. 154; no. 2; pp. 167 - 175
• Main Authors: Hu, Xiao-Hui, Ding, Li-Ya, Huang, Wei-Xue, Yang, Xian-Mei, Xie, Fang, Xu, Min, Yu, Long
• Format: Journal Article
• Language: English
• Published: England 01.08.2013
• Subjects: Catalytic Domain; Catechin - analogs & derivatives; Catechin - chemistry; Enzyme Inhibitors - chemistry; Escherichia coli; Gene Expression; Humans; Hydrogen Bonding; Molecular Docking Simulation; NADP - chemistry; NADP - genetics; NADP - metabolism; Recombinant Proteins - antagonists & inhibitors; Recombinant Proteins - chemistry; Recombinant Proteins - genetics; Recombinant Proteins - metabolism; Sugar Alcohol Dehydrogenases - antagonists & inhibitors; Sugar Alcohol Dehydrogenases - chemistry; Sugar Alcohol Dehydrogenases - genetics; Sugar Alcohol Dehydrogenases - metabolism; Surface Plasmon Resonance; mixed-type; SPR; inhibitor; DCXR; EGCG
• ISSN: 0021-924X; 1756-2651
• DOI: 10.1093/jb/mvt039

Dicarbonyl/l-xylulose reductase (DCXR), mainly catalysing the reduction of α-dicarbonyl compounds and l-xylulose, belongs to the short-chain dehydrogenase/reductase superfamily. Its enzyme activity can be inhibited by short-chain fatty acids. In this study, a novel DCXR inhibitor named (-)-epigallocatechin-3-gallate (EGCG) was reported. First, we overexpressed recombinant human DCXR in Escherichia coli, purified the enzyme by affinity chromatography and measured its activity. The inhibition effects of EGCG and its analogues on DCXR were determined subsequently, and EGCG showed the strongest inhibition with 50% inhibition concentration value of 78.8 μM. The surface plasmon resonance analysis also indicated that the equilibrium dissociation constant (KD) reached to 7.11 × 10(-8) M, which implied a high affinity between EGCG and DCXR. From enzyme kinetic analysis, EGCG acted as a mixed inhibitor against its forward and reverse substrates and the coenzyme, reduced nicotinamide adenine dinucleotide phosphate (NADPH). However, the inhibition is pH dependent. The molecular docking finally showed that EGCG formed several hydrogen bonds with the Thr190 residue of DCXR, and the model was further verified by site-directed mutagenesis. Therefore, EGCG is a potential inhibitor to human DCXR.