Promising Natural Compounds against Flavivirus Proteases: Citrus Flavonoids Hesperetin and Hesperidin
Abstract
:1. Introduction
2. Results and Discussion
2.1. Preparation of Flavivirus Proteases
2.2. Inhibition of DENV2, YFV, and WNV Proteases by HST and HSD
2.3. DENV2, YFV, and WNV NS2B/NS3pro–HST and HSD Interaction, as Determined by TFS
2.4. In Silico Studies of the Proteases–Flavonoids Interaction
3. Materials and Methods
3.1. Cloning of DENV2, YFV, and WNV Proteases
3.2. Heterologous Gene Expression and Purification of the Virus Proteases
3.3. Inhibition Assay of DENV2, YFV, and WNV NS2B/NS3pro
3.4. Characterisation of the Type of Inhibition and LE of HST and HSD
3.5. Intrinsic Tryptophan Fluorescence Spectroscopy (TFS)
3.6. Statistical Analysis
3.7. Molecular Dynamics and Computational Analysis
3.7.1. Ligand Parameterization
3.7.2. System Preparation
3.7.3. Simulation Setup
3.7.4. Molecular Dynamics Analysis
4. Conclusions
Supplementary Materials
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Acknowledgments
Conflicts of Interest
References
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Protease | Molecule | IC50 a | Inhibition Mode | pIC50 b | LE c | N d | Reference |
---|---|---|---|---|---|---|---|
DENV2 NS2B/NS3pro | HST | 4.7 ± 0.8 | noncompetitive | 5.33 | 0.34 | 22 | |
HSD | 55.7 ± 2.5 | noncompetitive | 4.25 | 0.14 | 43 | ||
YFV NS2B/NS3pro | HST | 2.0 ± 0.5 | noncompetitive | 5.69 | 0.36 | 22 | |
HSD | 67.7 ± 7.5 | noncompetitive | 4.17 | 0.13 | 43 | ||
WNV NS2B/NS3pro | HST | 4.3 ± 1.6 | noncompetitive | 5.37 | 0.34 | 22 | |
HSD | 50.7 ± 8.3 | noncompetitive | 4.29 | 0.14 | 43 | ||
ZIKV NS2B/NS3pro | HST | 12.6 ± 1.3 | noncompetitive | 4.90 | 0.31 | 22 | [11] |
HSD e | - | - | - | - | - | [11] |
Protease | Molecule | KD a | References |
---|---|---|---|
DENV2 NS2B/NS3pro | HST | 18.7 ± 2.7 | |
HSD | 20.9 ± 2.4 | ||
YFV NS2B/NS3pro | HST | 12.3 ± 2.2 | |
HSD | 29.5 ± 2.8 | ||
WNV NS2B/NS3pro | HST | 13.5 ± 2.0 | |
HSD | 19.5 ± 2.8 | ||
ZIKV NS2B/NS3pro | HST | 17.8 ± 2.9 | [11] |
HSD b | - | [11] |
Protein + Ligand | Replicate | Binding Energy [kcal/mol] | Representative Frames Cluster [%] |
---|---|---|---|
DENV2 NS2B/NS3pro–HST | 1 | −23.1 ± 3.6 | 68.9 |
2 | −24.1 ± 3.7 | 62.0 | |
YFV NS2B/NS3pro–HST | 1 | −20.0 ± 3.4 | 54.9 |
2 | −19.4 ± 2.7 | 64.8 | |
WNV NS2B/NS3pro–HST | 1 | −20.2 ± 3.4 | 72.7 |
2 | −28.3 ± 3.0 | 90.3 | |
DENV2 NS2B/NS3pro–HSD | 1 | −37.4 ± 7.2 | 95.3 |
2 | −38.6 ± 8.0 | 75.0 | |
YFV NS2B/NS3pro–HSD | 1 | −35.4 ± 5.2 | 36.9 |
2 | −37.6 ± 6.2 | 75.7 | |
WNV NS2B/NS3pro–HSD | 1 | −40.7 ± 4.7 | 63.1 |
2 | −37.8 ± 7.0 | 54.2 |
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Eberle, R.J.; Olivier, D.S.; Amaral, M.S.; Willbold, D.; Arni, R.K.; Coronado, M.A. Promising Natural Compounds against Flavivirus Proteases: Citrus Flavonoids Hesperetin and Hesperidin. Plants 2021, 10, 2183. https://doi.org/10.3390/plants10102183
Eberle RJ, Olivier DS, Amaral MS, Willbold D, Arni RK, Coronado MA. Promising Natural Compounds against Flavivirus Proteases: Citrus Flavonoids Hesperetin and Hesperidin. Plants. 2021; 10(10):2183. https://doi.org/10.3390/plants10102183
Chicago/Turabian StyleEberle, Raphael J., Danilo S. Olivier, Marcos S. Amaral, Dieter Willbold, Raghuvir K. Arni, and Monika A. Coronado. 2021. "Promising Natural Compounds against Flavivirus Proteases: Citrus Flavonoids Hesperetin and Hesperidin" Plants 10, no. 10: 2183. https://doi.org/10.3390/plants10102183