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

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Abstract

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 (Ka) at three temperatures (288, 298, and 308 K) were 7.940 × 107, 2.082 × 105 and 3.199 × 105 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: ΔHo (− 83.85 kJ mol−1), ΔGo (− 33.279 kJ mol−1), and ΔSo (− 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.

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Acknowledgements

This work was supported by the National Natural Science Foundation of China [Grant Numbers 31540012, 31470431, and 31670360], Guangdong Natural Science Foundation for Major cultivation project [Grant Number 2014A030308017], Shenzhen Science and Technology Innovation Committee Grants [Grant Numbers JSGG20160229120821300, JCYJ20150625103526744, CXZZ20150529165110750, JSGG20130411160539208, JCYJ20170302144535707, KQCX20140522111508785, CXZZ20150601110000604, and ZDSYS201506031617582], and Shenzhen special funds for Bio-industry development [Grant Number NYSW20140327010012].

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Author notes

  1. Manjunath D. Meti and Yang Xu have contributed equally as co-first authors in this paper.

Authors and Affiliations

  1. College of Life Sciences and Oceanography, Shenzhen Key Laboratory of Marine Bioresources and Ecology/Shenzhen Key Laboratory of Microbial Genetic Engineering, Shenzhen University, Shenzhen, 518060, Guangdong, People’s Republic of China

    Manjunath D. Meti, Yang Xu, Jiangfeng Xie, Yutao Chen, Zhibing Wu, Qingguo Han, Zhangli Hu & Hong Xu

  2. Key Laboratory of Optoelectronic Devices and Systems of Ministry of Education and Guangdong Province, College of Optoelectronic Engineering, Shenzhen University, Shenzhen, 518060, People’s Republic of China

    Manjunath D. Meti

  3. School of Science and Engineering, Chinese University of Hong Kong, Shenzhen, 518172, People’s Republic of China

    Yang Xu

  4. School of Science, Faculty of Health and Environmental Sciences, Auckland University of Technology, Auckland, 1142, New Zealand

    Yutao Chen

  5. School Medical Sciences, Faculty of Medicine, University of New South Wales, Sydney, NSW, 2052, Australia

    Johnson Liu

  6. School of Medicine, Shenzhen University, Shenzhen, 518060, People’s Republic of China

    Zhendan He

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  1. Manjunath D. Meti
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Correspondence to Hong Xu.

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Meti, M.D., Xu, Y., Xie, J. et al. Multi-spectroscopic studies on the interaction between traditional Chinese herb, helicid with pepsin. Mol Biol Rep 45, 1637–1646 (2018). https://doi.org/10.1007/s11033-018-4306-5

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