Multi-spectroscopic studies on the interaction between traditional Chinese herb, helicid with pepsin

• Published in: Molecular biology reports Vol. 45; no. 6; pp. 1637 - 1646
• Main Authors: Meti, Manjunath D., Xu, Yang, Xie, Jiangfeng, Chen, Yutao, Wu, Zhibing, Liu, Johnson, Han, Qingguo, He, Zhendan, Hu, Zhangli, Xu, Hong
• Format: Journal Article
• Language: English
• Published: Dordrecht Springer Netherlands 01-12-2018; Springer Nature B.V
• Subjects: Animal Anatomy; Animal Biochemistry; Benzaldehydes - chemistry; Binding Sites; Biomedical and Life Sciences; Circular Dichroism; Fluorescence; Herbal medicine; Histology; Hydrogen Bonding; Life Sciences; Medicine, Chinese Traditional; Molecular Docking Simulation - methods; Morphology; Original Article; Pepsin; Pepsin A - chemistry; Protein Binding - physiology; Protein Domains - physiology; Spectrometry, Fluorescence - methods; Spectrophotometry, Ultraviolet - methods; Temperature; Thermodynamics; Helicid; Pepsin; Multi-spectroscopic techniques; Molecular docking
• ISSN: 0301-4851; 1573-4978
• DOI: 10.1007/s11033-018-4306-5

Study on the binding properties of helicid by pepsin systematically using multi-spectroscopic techniques and molecular docking method, and these interactions comprise biological recognition at molecular level and backbone of biological significance in medicine concerned with the uses, effects, and modes of action of drugs. We investigated the mechanism of interaction between helicid and pepsin by using various spectroscopic techniques viz., fluorescence spectra, UV–Vis absorption spectra, circular dichroism (CD), 3D spectra, synchronous fluorescence spectra and molecular docking methods. The quenching mechanism associated with the helicid–pepsin interaction was determined by performing fluorescence measurements at different temperatures. From the experimental results show that helicid quenched the fluorescence intensity of pepsin via a combination of static and dynamic quenching process. The binding constants (
K
a
) at three temperatures (288, 298, and 308 K) were 7.940 × 10
7
, 2.082 × 10
5
and 3.199 × 10
5
L mol
−1
, respectively, and the number of binding sites (
n
) were 1.44, 1.14, and 1.18, respectively. The
n
value is close to unity, which means that there is only one independent class of binding site on pepsin for helicid. Thermodynamic parameters at 298 K were calculated as follows: Δ
H
o
(− 83.85 kJ mol
−1
), Δ
G
o
(− 33.279 kJ mol
−1
), and Δ
S
o
(− 169.72 J K
−1
mol
−1
). Based on thermodynamic analysis, the interaction of helicid with pepsin is driven by enthalpy, and Van der Waals’ forces and hydrogen bonds are the main forces between helicid and pepsin. A molecular docking study further confirmed the binding mode obtained by the experimental studies. The conformational changes in the structure of pepsin was confirmed by 3D fluorescence spectra and circular dichroism.