Title
Mechanism and Nature of Inhibition of Trypsin by Ligupurpuroside A, a Ku-Ding Tea Extract, Studied by Spectroscopic and Docking Methods
Author
Zhibing Wu, Liangliang Shen, Qingguo Han, Jun Lu, Haifeng Tang, Xu Xu, Hong Xu, Fengwen Huang, Jiangfeng Xie, Zhendan He, Zheling Zeng, Zhangli Hu
Year
2016
Journal
Food Biophysics
Abstract
Ligupurpuroside A is a glycoside extracted from Ku-Ding tea. As extracts from Ku-Ding tea exhibit anti-inflammatory property, we hypothesize that Ligupurpuroside A may be an active compound which inhibits trypsin activity during the anti-inflammatory process. The mechanism and nature of inhibition of trypsin by Ligupurpuroside A have been studied by multi-spectroscopic method, enzyme-activity assay and molecular docking. Enzyme activity assay reveals that Ligupurpuroside A significantly inhibits the activity of trypsin through a competitive manner with an IC50 value of 3.08 × 10−3 mol L−1. Fluorescence titration together with thermodynamic analysis indicate that a Ligupurpuroside A-trypsin complex is formed, and that hydrophobic force and hydrogen bonding are the main forces stabilizing the complex. UV-vis absorption, synchronous fluorescence and circular dichroism spectra show that the interaction between Ligupurpuroside A and trypsin induces conformational changes of trypsin with a decrease in the contents of α-helix and β-sheet. Finally, molecular docking further suggests that Ligupurpuroside A molecule binds within the active pocket of trypsin via hydrophobic force and hydrogen bond. Results from this study of the interaction of trypsin with its natural inhibitor should be useful to minimize the antinutritional effects and make full use of tea extracts in the food industry, and be also helpful to the design of the drugs for the diseases related to overexpression of trypsin.
Instrument
J-810
Keywords
Circular dichroism, Secondary structure, Ligand binding, Biochemistry, Food science