Mechanism and Nature of Inhibition of Trypsin by Ligupurpuroside A, a Ku-Ding Tea Extract, Studied by Spectroscopic and Docking Methods

• Published in: Food biophysics Vol. 12; no. 1; pp. 78 - 87
• Main Authors: Wu, Zhibing, Shen, Liangliang, Han, Qingguo, Lu, Jun, Tang, Haifeng, Xu, Xu, Xu, Hong, Huang, Fengwen, Xie, Jiangfeng, He, Zhendan, Zeng, Zheling, Hu, Zhangli
• Format: Journal Article
• Language: English
• Published: New York Springer US 01.03.2017; Springer Nature B.V
• Subjects: Analytical Chemistry; Biological and Medical Physics; Biophysics; Chemistry; Chemistry and Materials Science; Enzymatic activity; Fluorescence; Food Science; Molecular structure; Original Article; Proteases; Spectrum analysis; Tea; Spectroscopy; Docking; Trypsin; Ligupurpuroside A; Ku-Ding tea
• ISSN: 1557-1858; 1557-1866
• DOI: 10.1007/s11483-016-9465-0

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 IC 50 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.