Insights into Ergosterol Peroxide’s Trypanocidal Activity
Abstract
:1. Introduction
2. Materials and Methods
2.1. Protein Structure Preparation
2.2. Inverse Docking
2.3. Preliminary Surface Plasmon Resonance Experimental Procedures
2.4. CYP51 Protein Expression Trypanosoma cruzi and Candida albicans
2.5. Mushroom Strain
2.6. General Procedures
2.7. Isolation and Purification of Ergosterol Peroxide
2.8. Parasite Cultures
2.9. Carbonylation Activity in Extracellular Epimastigotes
2.10. Electron Microscopy Assay
2.11. Confocal Imaging
2.12. Transmission Electron Microscopy
2.13. Candida albicans Viability Assay
2.14. Analysis of Intermolecular SPR Interactions
3. Results
3.1. Ergosterol Peroxide Interacts with Tc Sterols and Does Not Target Tc Glucose Metabolism
3.2. CYP51 Interacts with Ergosterol Peroxide in Tc but Not in Candida albicans
3.3. Detection of Protein Carbonylation Assay
3.4. Detection of Enlarged Extracellular Vesicle Structures after Ergosterol Peroxide Treatment
3.5. Ergosterol Peroxide Disruption of the Cytoplasmic and Nuclear Membranes
3.6. Ergosterol Peroxide Disruption of the Golgi Apparatus and the Cytoplasmic and Nuclear Membranes
4. Discussion
5. Conclusions
Supplementary Materials
Author Contributions
Funding
Acknowledgments
Conflicts of Interest
References
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Trypanosoma spp. Protein Set | PDB Code | VINA Free energy of Binding (Kcal/moL) | ADT4.2 Free energy of Binding (Kcal/moL) | Calculated Ki (µM) |
---|---|---|---|---|
Sterol 14-alpha demethylase (CYP51) | 4CK9 | −11.1 | −12.21 | 0.002 |
Lanosterol 14 alpha demethylase | 3ZG2 | −10.9 | −11.29 | 0.013 |
Farnesyl transferase | 3WSB | −10.6 | −9.64 | 0.087 |
C-8 sterol demethylase model | 3bal * | −10.4 | −11.35 | 4.537 |
Squalene synthase (TcSQS) | 3WCA | −10.4 | −10.42 | 0.021 |
ABC b model | 4ayx * | −10.4 | −9.24 | 0.171 |
Phosphodiesterase | 3V93 | −10.1 | −10.32 | 28.66 |
Old Yellow enzyme (FMN oxidereductase) | 3ATY | −9.7 | −10.25 | 0.029 |
Glyceraldehyde 3-phosphate dehydrogenase | 1ML3 | −9.4 | −9.17 | 191.147 |
Trypanothione reductase | 2WBA | −9.2 | −10.06 | 44.723 |
Dihydroorotate dehydrogenase | 3W1R | −9.1 | −11.17 | 6.407 |
Chagasin | 3EZ1 | −8.4 | −7.59 | 2.9 |
Dihydrofolate reductase | 3KJS | −8.2 | −7.21 | 5060 |
Gluc-6-Pho Isomerase | 4QFH | −8 | −6.17 | 1830 |
Trans-sialidase | 3B69 | −7.9 | −9.34 | 0.145 |
Glutathione peroxidase | 3E0U | −7.6 | −7.5 | 3.24 |
Prostaglandin F synthase | 4GIE | −7.3 | −8.17 | 1.507 |
Sterol carrier protein 2 model | 3zbg * | −7.3 | −8.86 | 0.321 |
Cyclophilin | 1XQ7 | −7.1 | −7.88 | 1810 |
Lipoamide dehydrogenase | 2QAE | −7 | −5.22 | 9576.667 |
Cruzain | 4KLB | −6.9 | −7.58 | 2783.333 |
Fe-Superoxide dismutase | 2GPC | −6.8 | −6.52 | 16,176.667 |
Pyruvate kinase | 4KS0 | −6.7 | −9.59 | 91.953 |
Spermidine synthase | 4YV0 | −5.6 | −4.94 | 241.627 |
Dihydrofolate Reductase | 3IRN | −5.3 | −2.98 | 2,0330 |
Enolase | 4G7F | −3.9 | −7.3 | 102.843 |
Glucokinase | 5BRD | −3.4 | −2.57 | 310.043 |
Ribose 5-phosphate isomerase | 3K7O | −2.1 | −1.03 | 238.85 |
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Meza-Menchaca, T.; Ramos-Ligonio, A.; López-Monteon, A.; Vidal Limón, A.; Kaluzhskiy, L.A.; V. Shkel, T.; V. Strushkevich, N.; Jiménez-García, L.F.; Agredano Moreno, L.T.; Gallegos-García, V.; et al. Insights into Ergosterol Peroxide’s Trypanocidal Activity. Biomolecules 2019, 9, 484. https://doi.org/10.3390/biom9090484
Meza-Menchaca T, Ramos-Ligonio A, López-Monteon A, Vidal Limón A, Kaluzhskiy LA, V. Shkel T, V. Strushkevich N, Jiménez-García LF, Agredano Moreno LT, Gallegos-García V, et al. Insights into Ergosterol Peroxide’s Trypanocidal Activity. Biomolecules. 2019; 9(9):484. https://doi.org/10.3390/biom9090484
Chicago/Turabian StyleMeza-Menchaca, Thuluz, Angel Ramos-Ligonio, Aracely López-Monteon, Abraham Vidal Limón, Leonid A. Kaluzhskiy, Tatjana V. Shkel, Natallia V. Strushkevich, Luis Felipe Jiménez-García, Lourdes Teresa Agredano Moreno, Verónica Gallegos-García, and et al. 2019. "Insights into Ergosterol Peroxide’s Trypanocidal Activity" Biomolecules 9, no. 9: 484. https://doi.org/10.3390/biom9090484