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©The Author(s) 2020. Published by Baishideng Publishing Group Inc. All rights reserved.
World J Hepatol. Nov 27, 2020; 12(11): 976-992
Published online Nov 27, 2020. doi: 10.4254/wjh.v12.i11.976
Published online Nov 27, 2020. doi: 10.4254/wjh.v12.i11.976
Screening and identification of bioactive compounds from citrus against non-structural protein 3 protease of hepatitis C virus genotype 3a by fluorescence resonance energy transfer assay and mass spectrometry
Mahim Khan, Waqar Rauf, Fazal-e- Habib, Moazur Rahman, Mazhar Iqbal, Health Biotechnology Division, National Institute for Biotechnology and Genetic Engineering College, Pakistan Institute of Engineering and Applied Sciences (NIBGE-C, PIEAS), Faisalabad 38000, Punjab, Pakistan
Moazur Rahman, School of Biological Sciences, University of the Punjab, Lahore 54810, Punjab, Pakistan
Author contributions: Khan M was involved in this project from the design to execution and writing of this manuscript, optimized the expression and purification of hepatitis C virus non-structural protein 3 protease, extracted antioxidant compounds and tested their inhibition using the in vitro fluorescence resonance energy transfer assay, contributed to the LCMS/MS analysis to identify phenolics/flavonoids in plant extracts and evaluated their interaction using a modeling approach; Rauf W contributed to the design and execution of the experiments and LCMS/MS data analysis; Habib F performed the LCMS/MS analysis; Rahman M provided help with the purification of hepatitis C virus non-structural protein 3 and manuscript writing; Iqbal M conceived the idea, planned all of the experiments, and contributed to the data analysis, and manuscript writing.
Institutional review board statement: It is certified that the protocol of the study, screening and identification of bioactive compounds from citrus against non-structural protein 3 protease of hepatitis C virus genotype 3a by fluorescence resonance energy transfer assay and mass spectrometry was approved by the institutional review board.
Conflict-of-interest statement: There is no financial, commercial or other conflict of interest with any author.
Data sharing statement: All data generated or analyzed during this study are included in this manuscript and its supplementary material. Any additional information, if required, may be asked from the corresponding author (hamzamgondal@ gmail.com).
Open-Access: This article is an open-access article that was selected by an in-house editor and fully peer-reviewed by external reviewers. It is distributed in accordance with the Creative Commons Attribution NonCommercial (CC BY-NC 4.0) license, which permits others to distribute, remix, adapt, build upon this work non-commercially, and license their derivative works on different terms, provided the original work is properly cited and the use is non-commercial. See: http://creativecommons.org/Licenses/by-nc/4.0/
Corresponding author: Mazhar Iqbal, PhD, Professor, Health Biotechnology Division, National Institute for Biotechnology and Genetic Engineering College, Pakistan Institute of Engineering and Applied Sciences (NIBGE-C, PIEAS), Jhang Road, Faisalabad 38000, Punjab, Pakistan. hamzamgondal@gmail.com
Received: July 27, 2020
Peer-review started: July 27, 2020
First decision: August 22, 2020
Revised: September 3, 2020
Accepted: September 16, 2020
Article in press: September 16, 2020
Published online: November 27, 2020
Processing time: 119 Days and 23 Hours
Peer-review started: July 27, 2020
First decision: August 22, 2020
Revised: September 3, 2020
Accepted: September 16, 2020
Article in press: September 16, 2020
Published online: November 27, 2020
Processing time: 119 Days and 23 Hours
Core Tip
Core Tip: The manuscript describes the screening of active metabolites in citrus fruit extracts against hepatitis C virus genotype3a non-structural protein 3 (HCV-G3a NS3) protease. In this study, conditions have been optimized to get highly purified and functionally active protein HCV NS3. Further, fluorescence resonance energy transfer assay was used to screen the citrus extracts against NS3 protease. By using liquid chromatography coupled with tandem mass spectrometry/mass spectrometry analysis and bioinformatics modeling approaches, the observed activity of citrus extracts against HCV genotype3a NS3 protease was ascribed to hesperidin. Fluorescence resonance energy transfer assay confirmed the inhibitory potential of hesperidin against NS4A-NS3 protease domain with an half maximal inhibitory concentration value of 23.32 µmol/L.