Skip to main content

Advertisement

SpringerLink
Log in

Cloning, expression and dynamic simulation of TRYP6 from Leishmania major (MRHO/IR/75/ER)

  • Published:
Molecular Biology Reports Aims and scope Submit manuscript

Abstract

Leishmania, a digenetic protozoan parasite causes severe diseases in human and animals. Efficient evasion of toxic microbicidal molecules, such as reactive oxygen species and reactive nitrogen species is crucial for Leishmania to survive and replicate in the host cells. Tryparedoxin peroxidase, a member of peroxiredoxins family, is vital for parasite survival in the presence of antioxidant, hence it is one of the most important molecules in Leishmania viability and then, it may be an appropriate goal for challenging against leishmaniasis. After cloning and sub-cloning of TRYP6 from Leishmania major (MRHO/IR/75/ER), homology modeling of the LmTRYP6 was proposed to predict some functional property of this protein. The refined model showed that the core structure consists of a seven β stranded β-sheet and five α helices which are organized as a central 7-stranded β2-β1-β5-β4-β3-β6-β7 surrounded by 2-stranded β-hairpin, α helices A and D on one side, and α helices B, C and E on the other side. The peroxidatic active site is located in a pocket formed by the residue Pro45, Met46, Thr49, Val51, Cys52, Arg128, Met147 and Pro 148. The catalytic Cys52, located in the first turn of helix αB, is in van der Waals with a Pro45, a Thr49 and an Arg128 that are absolutely conserved in all known Prx sequences. In this study, an attractive molecular target was studied. These results might be used in designing of drugs to fight an important human pathogen.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5
Fig. 6
Fig. 7

Similar content being viewed by others

References

  1. Channon JY, Roberts MB, Blackwell JM (1984) A study of the differential respiratory burst activity elicited by promastigotes and amastigotes of Leishmania donovani in murine resident peritoneal macrophages. Immunology 53:345–355

    PubMed  CAS  Google Scholar 

  2. Murray HW (1982) Cell-mediated immune response in experimental visceral leishmaniasis II. Oxygen-dependent killing of intracellular Leishmania donovani amastigotes. J Immunol 129:351–357

    PubMed  CAS  Google Scholar 

  3. Muller S, Liebau E, Walter RD, Krauth-Siegel RL (2003) Thiol based redox metabolism of protozoan parasites. Trends Parasitol 19:320–328

    Article  PubMed  CAS  Google Scholar 

  4. Tovar J, Cunningham ML, Smith AC, Croft SL, Fairlamb AH (1998) Down-regulation of Leishmania donovani trypanothione reductase by heterologous expression of a trans-dominant mutant homologue: effect on parasite intracellular survival. Proc Natl Acad Sci USA 95:5311–5316

    Article  PubMed  CAS  Google Scholar 

  5. Krauth-Siegel RL, Meiering SK, Schmidt H (2003) The parasite-specific trypanothione metabolism of trypanosoma and Leishmania. Biol Chem 384:539–549

    Article  PubMed  CAS  Google Scholar 

  6. Dumas C, Ouellette M, Tovar J, Cunningham ML, Fairlamb AH, Tamar S, Olivier M, Papadopoulou B (1997) Disruption of the trypanothione reductase gene of Leishmania decreases its ability to survive oxidative stress in macrophages. EMBO J 16:2590–2598

    Article  PubMed  CAS  Google Scholar 

  7. Schmidt A, Krauth-Siegel RL (2002) Enzymes of the trypanothione metabolism as targets for antitrypanosomal drug development. Curr Top Med Chem 2:1239–1259

    Article  PubMed  CAS  Google Scholar 

  8. Levick MP, Tetaud E, Fairlamb AH, Blackwell JM (1998) Identification and characterisation of a functional peroxidoxin from Leishmania major. Mol Biochem Parasitol 96:125–137

    Article  PubMed  CAS  Google Scholar 

  9. Zarley JH, Britigan BE, Wilson ME (1991) Hydrogen peroxide-mediated toxicity for Leishmania donovani chagasi promastigotes. Role of hydroxyl radical and protection by heat shock. J Clin Invest 88:1511–1521

    Article  PubMed  CAS  Google Scholar 

  10. Wilson ME, Andersen KA, Britigan BE (1994) Response of Leishmania chagasi promastigotes to oxidant stress. Infect Immun 62:5133–5141

    PubMed  CAS  Google Scholar 

  11. Lin YC, Hsu JY, Chiang SC, Lee ST (2005) Distinct overexpression of cytosolic and mitochondrial tryparedoxin peroxidases results in preferential detoxification of different oxidants in arsenite-resistant Leishmania amazonensis with and without DNA amplification. Mol Biochem Parasitol 142:66–75

    Article  PubMed  CAS  Google Scholar 

  12. Walker J, Acestor N, Gongora R, Quadroni M, Segura I, Fasel N, Saravia NG (2006) Comparative protein profiling identifies elongation factor-1beta and tryparedoxin peroxidase as factors associated with metastasis in Leishmania guyanensis. Mol Biochem Parasitol 145:254–264

    Article  PubMed  CAS  Google Scholar 

  13. Harder S, Bente M, Isermann K, Bruchhaus I (2006) Expression of a mitochondrial peroxiredoxin prevents programmed cell death in Leishmania donovani. Eukaryot Cell 5(5):861–870

    Article  PubMed  CAS  Google Scholar 

  14. Chen L, Xie QW, Nathan C (1998) Alkyl hydroperoxide reductase subunit C (AhpC) protects bacterial and human cells against reactive nitrogen intermediates. Mol Cell 1(6):795–805

    Article  PubMed  CAS  Google Scholar 

  15. Bryk R, Griffin P, Nathan C (2000) Peroxynitrite reductase activity of bacterial peroxiredoxins. Nature 407(6801):211–215

    Article  PubMed  CAS  Google Scholar 

  16. Castro H, Budde H, Flohé L, Hofmann B, Lünsdorf H, Wissing J, Tomás AM (2002) Specificity and kinetics of a mitochondrial peroxiredoxin of Leishmania infantum. Free Radic Biol Med 33(11):1563–1574

    Article  PubMed  CAS  Google Scholar 

  17. Castro H, Sousa C, Santos M, Cordeiro-da-Silva A, Flohé L, Tomás AM (2002) Complementary antioxidant defense by cytoplasmic and mitochondrial peroxiredoxins in Leishmania infantum. Free Radic Biol Med 33(11):1552–1562

    Article  PubMed  CAS  Google Scholar 

  18. Flohé L, Budde H, Bruns K, Castro H, Clos J, Hofmann B, Kansal-Kalavar S, Krumme D, Menge U, Plank-Schumacher K, Sztajer H, Wissing J, Wylegalla C, Hecht HJ (2002) Tryparedoxin peroxidase of Leishmania donovani: molecular cloning, heterologous expression, specificity, and catalytic mechanism. Arch Biochem Biophys 397(2):324–335

    Article  PubMed  Google Scholar 

  19. Barr SD, Gedamu L (2001) Cloning and characterization of three differentially expressed peroxidoxin genes from Leishmania chagasi. Evidence for an enzymatic detoxification of hydroxyl radicals. J Biol Chem 276(36):34279–34287

    Article  PubMed  CAS  Google Scholar 

  20. Khalil EAG, El Hassan Zijlstra EE, Mukhtat MM, Ghalib HW, Musa B, Ibrahim ME, Kamil A, Elsheikh M, Modabber F (2000) Autoclaved L. major vaccine for prevention of visceral leishmaniasis: a randomised, double-blind, BCG-controlled trial in Sudan. Lancet 356:1565–1569

    Article  PubMed  CAS  Google Scholar 

  21. Nadim A, Javadian E (1998) Leishmanization in Iran. In: Walton B, Wijeyaretne PM, Modabber F (eds) Research on strategies for the control of Leishmaniasis. International Development Research Center, Ottawa, pp 336–369

    Google Scholar 

  22. Sharifi I, FeKri AR, Aflatonian MR, Khamesipour A, Nadim A, Mousavi MR, Momeni AZ, Dowlati Y, Godal T, Zicker F, Smith PG, Modabber F (1998) Randomised vaccine trial of single dose of killed Leishmania major plus BCG against anthroponotic cutaneous leishmaniasis in Bam, Iran. Lancet 351(9115):1540–1543

    Article  PubMed  CAS  Google Scholar 

  23. Momeni AZ, Jalayer T, Emamjomeh M, Khamesipour A, Zicker F, Ghassemi RL, Dowlati Y, Sharifi I, Aminjavaheri M, Shafiei A, Alimohammadian MH, Hashemi-Fesharki R, Nasseri K, Godal T, Smith PG, Modabber F (1997) A randomised, double-blind, controlled trial of a killed L. major vaccine plus BCG against zoonotic cutaneous leishmaniasis in Iran. Vaccine 17(5):466–472

    Article  Google Scholar 

  24. Bahar K, Dowlati Y, Shidani B, Alimohammadian MH, Khamesipour A, Ehsasi S, Hashemi-Fesharki R, Ale-Agha S, Modabber F (1996) Comparative safety and immunogenicity trial of two killed Leishmania major with or without BCG in human volunteers. Clin Dermatol 14(5):489–495

    Article  PubMed  CAS  Google Scholar 

  25. Khamesipour A, Dowlati Y, Asilian A, Hashemi-Fesharki R, Javadi A, Noazin S, Modabber F (2005) Leishmanization: use of an old method for evaluation of candidate vaccines against leishmaniasis. Vaccine 23(28):3642–3648

    Article  PubMed  CAS  Google Scholar 

  26. Hendricks LD, Wood DE, Hajduk ME (1978) Haemoflagellates commercially available liquid media for rapid cultivation. Parasitology 76:309–316

    Article  PubMed  CAS  Google Scholar 

  27. Ozbilgin A, Tas S, Atambay M, Alkan Z, O¨ zbel Y (1995) Comparison of liquid and biphasic media in in vitro cultivation of Leishmania infantum promastigotes. Acta Parasitol Turc 19:1–5

    Google Scholar 

  28. Castro H, Sousa C, Novais M, Santos M, Budde H, Cordeiro-da-Silva A, Flohe L, Tomas AM (2004) Two linked genes of Leishmania infantum encode tryparedoxins localised to cytosol and Mitochondrion. Mol Biochem Parasitol 136:137–147

    Article  PubMed  CAS  Google Scholar 

  29. Eisenberger CL, Jaffe CL (1999) Leishmania: identification of old world species using a permissively primed intergenic polymorphic-polymerase chain reaction. Exp Parasitol 91:70–77

    Article  PubMed  CAS  Google Scholar 

  30. Altschul SF, Madden TL, Schaffer AA, Zhang J, Zhang Z, Miller W, Lipman DJ (1997) Gapped BLAST and PSI-BLAST: a new generation of protein database search programs. Nucleic Acids Res 25:3389–3402

    Article  PubMed  CAS  Google Scholar 

  31. Altschul SF, Wootton JC, Gertz EM, Agarwala R, Morgulis A, Schaffer AA, Yu Y (2005) Protein database searches using compositionally adjusted substitution matrices. FEBS J 272:5101–5109

    Article  PubMed  CAS  Google Scholar 

  32. Kang GY, Park EH, Lim CJ (2008) Molecular cloning, characterization and regulation of a peroxiredoxin gene from Schizosaccharomyces pombe. Mol Biol Rep 35(3):387–395

    Article  PubMed  CAS  Google Scholar 

  33. Qiu L, Ma Z, Jiang S, Wang W, Zhou F, Huang J, Li J, Yang Q (2010) Molecular cloning and mRNA expression of peroxiredoxin gene in black tiger shrimp (Penaeus monodon). Mol Biol Rep 37(6):2821–2827

    Article  PubMed  CAS  Google Scholar 

  34. Finkemeier I, Goodman M, Lamkemeyer P, Kandlbinder A, Sweetlove LJ, Dietz KJ (2005) The mitochondrial type II peroxiredoxin F is essential for redox homeostasis and root growth of Arabidopsis thaliana under stress. J Biol Chem 280(13):12168–12180

    Article  PubMed  CAS  Google Scholar 

  35. Cong M, Ni D, Song L, Wang L, Zhao J, Qiu L, Li L (2009) Molecular cloning, characterization and mRNA expression of peroxiredoxin in Zhikong scallop Chlamys farreri. Mol Biol Rep 36(6):1451–1459

    Article  PubMed  CAS  Google Scholar 

  36. Peng Y, Yang PH, Guo Y, Ng SS, Liu J, Fung PC, Tay D, Ge J, He ML, Kung HF, Lin MC (2004) Catalase and peroxiredoxin 5 protect Xenopus embryos against alcohol-induced ocular anomalies. Invest Ophthalmol Vis Sci 45(1):23–29

    Article  PubMed  Google Scholar 

  37. Hofmann B, Hecht HJ, Flohé L (2002) Peroxiredoxins. Biol Chem 383:347–364

    Article  PubMed  CAS  Google Scholar 

  38. Trivelli X, Krimm I, Ebel C, Verdoucq L, Prouzet-Mauléon V, Chartier Y, Tsan P, Lauquin G, Meyer Y, Lancelin JM (2003) Characterization of the yeast peroxiredoxin Ahp1 in its reduced active and over oxidized inactive forms using NMR. Biochemistry 42:14139–14149

    Article  PubMed  CAS  Google Scholar 

  39. Flohe L, Harris JR (2007) Peroxiredoxin systems, subcellular biochemistry. Springer, Germany

    Google Scholar 

  40. Seo MS, Kang SW, Kim K, Baines IC, Lee TH, Rhee SG (2000) Identification of a new type of mammalian peroxiredoxin that forms an intramolecular disulfide as a reaction intermediate. J Biol Chem 275:20346–20354

    Article  PubMed  CAS  Google Scholar 

  41. Rhee SG, Chae HZ, Kim K (2005) Peroxiredoxins: a historical overview and speculative preview of novel mechanisms and emerging concepts in cell signaling. Free Radic Biol Med 38:1543–1552

    Article  PubMed  CAS  Google Scholar 

  42. Li H, Robertson AD, Jensen JH (2005) Very fast empirical prediction and rationalization of protein pKa values. Proteins 61:704–721

    Article  PubMed  CAS  Google Scholar 

  43. IMHME (2009) Official report of Leishmania cases in Iran. Iran Ministry of Health & Medical Education (IMHME)

  44. Mittal K, Rai S, Ravinder A, Gupta S, Sundar S, Goyal N (2007) Characterization of natural antimony resistance in Leishmania donovani isolates. Am J Trop Med Hyg 76:681–688

    PubMed  CAS  Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Department of Parasitology and Mycology, Shahid Sadoughi University of Medical Sciences, 8916188/35, Yazd, Iran

    G. Eslami

  2. Laboratoire de Biochimie et de Génie Enzymatique des Lipases, École Nationale d’Ingénieurs de Sfax, Route Soukra Km 3.5 B.P W, 1173, Sfax, Tunisia

    F. Frikha

  3. Department of Genetics and Molecular Biology, Faculty of Medicine, Isfahan University of Medical Sciences, Isfahan, Iran

    R. Salehi

  4. Center for Research and Training in Skin Disease and Leprosy, Tehran University of Medical Sciences, 415 Taleghani Avenue, 1416613675, Tehran, Iran

    A. Khamesipour & M. A. Nilforoushzadeh

  5. Department of Parasitology and Mycology, Faculty of Medicine, Isfahan University of Medical Sciences, Isfahan, Iran

    H. Hejazi

  6. Skin Diseases and Leishmaniasis Research Center, Isfahan University of Medical Sciences, Isfahan, Iran

    M. A. Nilforoushzadeh

Authors
  1. G. Eslami
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. F. Frikha
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. R. Salehi
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. A. Khamesipour
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. H. Hejazi
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. M. A. Nilforoushzadeh
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to G. Eslami.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Eslami, G., Frikha, F., Salehi, R. et al. Cloning, expression and dynamic simulation of TRYP6 from Leishmania major (MRHO/IR/75/ER). Mol Biol Rep 38, 3765–3776 (2011). https://doi.org/10.1007/s11033-010-0492-5

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s11033-010-0492-5

Keywords

Search

Navigation