Trypanothione Reductase Gene Mutations in Meglumine Antimoniate Resistant Isolates from Cutaneous Leishmaniasis Patients Using Molecular Dynamics Method
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Abstract
Background: In this study, mutations in the tripanothion reductase of Leishmania tropica isolated from Iran was investigated using sequencing and simulation of the enzyme by the molecular dynamic method.
Methods: Fifteen susceptible and 15 clinical resistant L. tropica specimens were collected from skin lesions from different regions of Iran in 2017. After DNA extraction, trypanothione reductase (TRYR or TPR), gene fragment was amplified using PCR and sequencing methods. In the case of structural mutations, the components were simulated by molecular dynamics using the GROMACS software.
Results: Some structural mutations were observed in 9 amino acids surrounding the active site of the TRYR gene of L. tropica with three-dimensional trypanothione reductase alteration.
Conclusion: Change in the active site of TRYR of L. tropica, could probably contribute to the development of resistant L. tropica to glucantime. Because of the likely occurrence of mutations in glucantime as well as the ease of development of L. tropica resistant populations, more samples are needed to demonstrate the relationship between mutations in this enzyme and clinical resistance to glucantime. On the other hand, it is recommended that enzymatic studies be performed to confirm the role of mutation in the function and expression of trypanothione reductase in glucantime resistant and susceptible populations.
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2. Holakouie-Naieni K, Mostafavi E, Boloorani AD, et al. Spatial modeling of cutaneous leishmaniasis in Iran from 1983 to 2013. Acta Trop. 2017;166:67-73.
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11. Grondin K, Haimeur A, Mukhopadhyay R, et al. Co-amplification of the gamma-glutamylcysteine synthetase gene gsh1 and of the ABC transporter gene pgpA in arsenite-resistant Leishmania tarentolae. EMBO J. 1997;16(11):3057-65.
12. Zhou Y, Messier N, Ouellette M, et al. Leishmania major LmACR2 is a pentavalent antimony reductase that confers sensitivity to the drug pentostam. J Biol Chem. 2004;279(36):37445-51.
13. Baiocco P, Colotti G, Franceschini S, et al. Molecu-lar basis of antimony treatment in leishmaniasis. J Med Chem. 2009;52(8):2603-12.
14. Yan S, Wong IL, Chow LM, et al. Rapid reduction of pentavalent antimony by trypanothione: potential relevance to antimonial activation. Chem Commun (Camb). 2003;(2):266-7.
15. Yan S, Li F, Ding K, et al. Reduction of pentavalent antimony by trypanothione and formation of a bina-ry and ternary complex of antimony(III) and trypa-nothione. J Biol Inorg Chem. 2003;8(6):689-97.
16. Ferreira Cdos S, Martins PS, Demicheli C, et al. Thiol-induced reduction of antimony (V) into anti-mony (III): a comparative study with trypanothione, cysteinyl-glycine, cysteine and glutathione. Biomet-als. 2003;16(3):441-6.
17. Vickers TJ, Fairlamb AH. Trypanothione S-transferase activity in a trypanosomatid ribosomal elongation factor 1B. J Biol Chem. 2004;279(26):27246-56.
18. Denton H, McGregor JC, Coombs GH. Reduc-tion of anti-leishmanial pentavalent antimonial drugs by a parasite-specific thiol-dependent reductase, TDR1. Biochem J. 2004;381(Pt 2):405-12.
19. Hadighi R, Mohebali M, Boucher P, et al. Unre-sponsiveness to glucantime treatment in Iranian cu-taneous leishmaniasis due to drug-resistant Leishma-nia tropica parasites. PLoS Med. 2006;3(5):e162.
20. Kazemi-Rad E, Mohebali M, Khadem-Erfan MB, et al. Overexpression of ubiquitin and amino acid permease genes in association with antimony re-sistance in Leishmania tropica field isolates. Korean J Parasitol. 2013;51(4):413-9.
21. Kazemi-Rad E, Mohebali M, Khadem-Erfan MB, et al. Identification of antimony resistance markers in Leishmania tropica field isolates through a cDNA-AFLP approach. Exp Parasitol. 2013;135(2):344-9.
22. Mohebali M, Kazemirad E, Hajjaran H, et al. Gene expression analysis of antimony resistance in Leish-mania tropica using quantitative real-time PCR fo-cused on genes involved in trypanothione metabo-lism and drug transport. Arch Dermatol Res. 2019;311(1):9-17.
23. Erfan MBK, Mohebali M, Kazemi-Rad E, et al. Downregulation of calcineurin gene is associated with Glucantime® resiatance in Leishmania infantum. Iran J Parasitol. 2013;8(3):359-66.
24. Hajjaran H, Azarian B, Mohebali M, et al. Compara-tive proteomics study on meglumine antimoniate sensitive and resistant Leishmania tropica isolated from Iranian anthroponotic cutaneous leishmaniasis pa-tients. East Mediterr Health J. 2012;18(2):165-71.
25. Cunningham ML, Zvelebil MJ, Fairlamb AH. Mechanism of inhibition of trypanothione reduc-tase and glutathione reductase by trivalent organic arsenicals. Eur J Biochem. 1994; 221(1):285-95.
26. Torres DC, Ribeiro-Alves M, Romero GA, et al. Assessment of drug resistance related genes as candidate markers for treatment outcome predic-tion of cutaneous leishmaniasis in Brazil. Acta Trop. 2013;126(2):132-41.
2. Holakouie-Naieni K, Mostafavi E, Boloorani AD, et al. Spatial modeling of cutaneous leishmaniasis in Iran from 1983 to 2013. Acta Trop. 2017;166:67-73.
3. Heydarpour F, Sari AA, Mohebali M, et al. Inci-dence and disability-adjusted life years (Dalys) at-tributable to leishmaniasis in Iran, 2013. Ethiop J Health Sci. 2016;26(4):381-8.
4. Bottger EC, Springer B. Tuberculosis: drug re-sistance, fitness, and strategies for global control. Eur J Pediatr. 2008;167(2):141-8.
5. Padron-Nieves M, Ponte-Sucre A, Diaz E. Drug resistance in Leishmania parasites: Consequences, molecular mechanisms and possible treatments. Heidelberg New York Dordrecht London: Spring-er-Verlag Wien; 2013. 462.
6. Croft SL. Monitoring drug resistance in leishmania-sis. Trop Med Int Health. 2001; 6(11):899-905.
7. Blundell TL, Sibanda BL, Montalvao RW, et al. Structural biology and bioinformatics in drug de-sign: opportunities and challenges for target identifi-cation and lead discovery. Philos Trans R Soc Lond B Biol Sci. 2006; 361(1467):413-23.
8. Frezard F, Demicheli C, Ribeiro RR. Pentavalent antimonials: new perspectives for old drugs. Mole-cules. 2009; 14(7):2317-36.
9. Wyllie S, Fairlamb AH. Differential toxicity of an-timonial compounds and their effects on glutathi-one homeostasis in a human leukaemia monocyte cell line. Biochem Pharmacol. 2006; 71(3):257-67.
10. Saha P, Mukhopadhyay D, Chatterjee M. Im-munomodulation by chemotherapeutic agents against leishmaniasis. Int Immunopharmacol. 2011;11(11):1668-79.
11. Grondin K, Haimeur A, Mukhopadhyay R, et al. Co-amplification of the gamma-glutamylcysteine synthetase gene gsh1 and of the ABC transporter gene pgpA in arsenite-resistant Leishmania tarentolae. EMBO J. 1997;16(11):3057-65.
12. Zhou Y, Messier N, Ouellette M, et al. Leishmania major LmACR2 is a pentavalent antimony reductase that confers sensitivity to the drug pentostam. J Biol Chem. 2004;279(36):37445-51.
13. Baiocco P, Colotti G, Franceschini S, et al. Molecu-lar basis of antimony treatment in leishmaniasis. J Med Chem. 2009;52(8):2603-12.
14. Yan S, Wong IL, Chow LM, et al. Rapid reduction of pentavalent antimony by trypanothione: potential relevance to antimonial activation. Chem Commun (Camb). 2003;(2):266-7.
15. Yan S, Li F, Ding K, et al. Reduction of pentavalent antimony by trypanothione and formation of a bina-ry and ternary complex of antimony(III) and trypa-nothione. J Biol Inorg Chem. 2003;8(6):689-97.
16. Ferreira Cdos S, Martins PS, Demicheli C, et al. Thiol-induced reduction of antimony (V) into anti-mony (III): a comparative study with trypanothione, cysteinyl-glycine, cysteine and glutathione. Biomet-als. 2003;16(3):441-6.
17. Vickers TJ, Fairlamb AH. Trypanothione S-transferase activity in a trypanosomatid ribosomal elongation factor 1B. J Biol Chem. 2004;279(26):27246-56.
18. Denton H, McGregor JC, Coombs GH. Reduc-tion of anti-leishmanial pentavalent antimonial drugs by a parasite-specific thiol-dependent reductase, TDR1. Biochem J. 2004;381(Pt 2):405-12.
19. Hadighi R, Mohebali M, Boucher P, et al. Unre-sponsiveness to glucantime treatment in Iranian cu-taneous leishmaniasis due to drug-resistant Leishma-nia tropica parasites. PLoS Med. 2006;3(5):e162.
20. Kazemi-Rad E, Mohebali M, Khadem-Erfan MB, et al. Overexpression of ubiquitin and amino acid permease genes in association with antimony re-sistance in Leishmania tropica field isolates. Korean J Parasitol. 2013;51(4):413-9.
21. Kazemi-Rad E, Mohebali M, Khadem-Erfan MB, et al. Identification of antimony resistance markers in Leishmania tropica field isolates through a cDNA-AFLP approach. Exp Parasitol. 2013;135(2):344-9.
22. Mohebali M, Kazemirad E, Hajjaran H, et al. Gene expression analysis of antimony resistance in Leish-mania tropica using quantitative real-time PCR fo-cused on genes involved in trypanothione metabo-lism and drug transport. Arch Dermatol Res. 2019;311(1):9-17.
23. Erfan MBK, Mohebali M, Kazemi-Rad E, et al. Downregulation of calcineurin gene is associated with Glucantime® resiatance in Leishmania infantum. Iran J Parasitol. 2013;8(3):359-66.
24. Hajjaran H, Azarian B, Mohebali M, et al. Compara-tive proteomics study on meglumine antimoniate sensitive and resistant Leishmania tropica isolated from Iranian anthroponotic cutaneous leishmaniasis pa-tients. East Mediterr Health J. 2012;18(2):165-71.
25. Cunningham ML, Zvelebil MJ, Fairlamb AH. Mechanism of inhibition of trypanothione reduc-tase and glutathione reductase by trivalent organic arsenicals. Eur J Biochem. 1994; 221(1):285-95.
26. Torres DC, Ribeiro-Alves M, Romero GA, et al. Assessment of drug resistance related genes as candidate markers for treatment outcome predic-tion of cutaneous leishmaniasis in Brazil. Acta Trop. 2013;126(2):132-41.
| Files | ||
| Issue | Vol 15 No 4 (2020) | |
| Section | Original Article(s) | |
| DOI | https://doi.org/10.18502/ijpa.v15i4.4856 | |
| Keywords | ||
| Leishmania tropica Glucantime Clinical resistance Tripanothion reductase | ||
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How to Cite
1.
FOZONGARI F, DALIMI A, ARAB SS, BEHMANESH M, KHAMMARI A. Trypanothione Reductase Gene Mutations in Meglumine Antimoniate Resistant Isolates from Cutaneous Leishmaniasis Patients Using Molecular Dynamics Method. Iran J Parasitol. 2020;15(4):511-520.
