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Molecular cloning and enzymatic characterization of a class mu glutathione S-transferase of Paragonimus westermani

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Abstract

Glutathione S-transferase (GST) is a component of a second line of defense against bioreactive radicals derived from host immune attack. Paragonimus westermani causes acute or chronic lung diseases in mammals. A cDNA clone, PwGST#11, of adult P. westermani produced in the present study was 748 bp long and encoded an open reading frame of 217 amino acids with a starting methionine. The molecular mass of this putative polypeptide, Pw26GST, was estimated to be 25.1 kDa with an isoelectric point of 5.7. Pw26GST was homologous with the 26-kDa GSTs of trematodes and vertebrates. Nine of the ten amino acid residues lining the glutathione-binding pocket were conserved. Putative Pw26GST polypeptide was clustered with 26-kDa GSTs of trematodes belonging to the class μ. Recombinant Pw26GST protein generated bacterially, revealed GST enzyme activity toward an universal and class μ-specific substrates. Mouse antisera to recombinant Pw26GST protein recognized native 26-kDa GST of P. westermani but not the GSTs of any other trematodes. Collectively, Pw26GST was found to be a member of class μ GSTs of P. westermani.

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References

  • Balloul JM, Sondermeyer P, Capron M, Grzych MM, Pierce RJ, Carvallo D, Lecocq JP, Capron A (1987) Molecular cloning of a protective antigen of Schistosome. Nature 326:149–153

    Article  PubMed  CAS  Google Scholar 

  • Blair D, Xu ZB, Agatsuma T (1999) Paragonimiasis and the genus Paragonimus. Adv Parasitol 42:113–222

    Article  PubMed  CAS  Google Scholar 

  • Boulanger D, Warter A, Trottein F, Mauny F, Bremond P, Audibert F, Couret D, Kadri S, Godin C, Sellin E, Pierce RJ, Lecocq JP, Sellin B, Capron A (1995) Vaccination of patas monkeys experimentally infected with Schistosoma haematobium using a recombinant glutathione S-transferase cloned from S. mansoni. Parasite Immunol 17:361–369

    PubMed  CAS  Google Scholar 

  • Brophy PM, Pritchard DI (1994) Parasitic helminth glutathione S-transferase: an update on their potential as targets for immuno- and chemotherapy. Exp Parasitol 79:89–96

    Article  PubMed  CAS  Google Scholar 

  • Brophy PM, Papadopoulos A, Touraki M, Coles B, Korting W, Barrett J (1989) Purification of cytosolic glutathione transferase from Schistocephalus solidus (plerocercoid) interaction with anthelmintics and products of lipid peroxidation. Mol Biochem Parasitol 36:187–195

    Article  PubMed  CAS  Google Scholar 

  • Chung YB, Song CY, Lee HS, Kong Y, Cho SY (1991) Purification and characterization of a Cu, Zn-superoxide dismutase from adult Paragonimus westermani. Kisengchunghak Chapji 29:259–266

    CAS  Google Scholar 

  • Chung YB, Lee HS, Song CY, Cho SY (1992) Activities of scavenging enzymes of oxygen radicals in early maturation stages of Paragonimus westermani. Kisengchunghak Chapji 30:355–358

    CAS  Google Scholar 

  • Erhardt J, Dirr HW (1995) Native dimer stabilizes the subunit tertiary structure of porcine class pi glutathione S-transferase. Eur J Biochem 230:614–620

    Article  PubMed  CAS  Google Scholar 

  • Habig WH, Pabst MJ, Jacoby WB (1974) Glutathione S-transferase: first enzymatic step in mercapturic acid formation. J Biol Chem 249:7130–7139

    PubMed  CAS  Google Scholar 

  • Hayes JD, Pulford DJ (1995) The glutathione S-transferase supergene family: regulation of GST and the contribution of the isoenzymes to cancer chemoprotection and drug resistance. Crit Rev Biochem Mol Biol 30:445–600

    PubMed  CAS  Google Scholar 

  • Hong SJ, Kang SY, Chung YB, Chung MH, Oh Y-J, Kang I, Bahk YY, Kong Y, Cho SY (2000) Paragonimus westermani: a cytosolic glutathione S-transferase of a σ-class in adult stage. Exp Parasitol 94:180–189

    Article  PubMed  CAS  Google Scholar 

  • Hong SJ, Lee JY, Lee DH, Sohn WM, Cho SY (2001) Molecular cloning and characterization of a mu-class glutathione S-transferase from Clonorchis sinensis. Mol Biochem Parasitol 115:69–75

    Article  PubMed  CAS  Google Scholar 

  • Ji X, Zhang P, Armstrong RN, Gilliland GL (1992) The three-dimensional structure of a glutathione S-transferase from the mu gene class. Structural analysis of the binary complex of isozyme 3-3 and glutathione at 2.2 Å resolution. Biochemistry 31:10169–10184

    Article  PubMed  CAS  Google Scholar 

  • Liebau E, Schonberger OL, Walter RD, Henkle-Duhrsen K (1994a) Molecular cloning and expression of a cDNA encoding glutathione S-transferase from Ascaris suum. Mol Biochem Parasitol 63:167–170

    Article  PubMed  CAS  Google Scholar 

  • Liebau E, Walter RD, Henkle-Duhrsen K (1994b) Isolation, sequence and expression of an Onchocerca volvulus glutathione S-transferase cDNA. Mol Biochem Parasitol 63:305–309

    Article  PubMed  CAS  Google Scholar 

  • Lim K, Ho JX, Keeling K, Gilliland GL, Ji X, Ruker F, Carter DC (1994) Three-dimensional structure of Schistosoma japonicum glutathione S-transferase fused with a six amino acid conserved neutralizing epitope of gp41 from HIV. Protein Sci 3:2233–2244

    Article  PubMed  CAS  Google Scholar 

  • Liu LF, Tam MF (1991) Nucleotide sequence of a class mu glutathione S-transferase from chicken liver. Biochim Biophys Acta 1090:343–344

    PubMed  CAS  Google Scholar 

  • Mannervik B, Alin P, Guthenberg C, Jensson H, Tahir MK, Warholm M, Jornvall H (1985) Identification of three classes cytosolic glutathione transferase common to several mammalian species: correlation between structural data and enzymatic properties. Proc Natl Acad Sci U S A 82:7202–7206

    Article  PubMed  CAS  Google Scholar 

  • McNair AT, Dissous C, Duvaux-Miret O, Capron A (1993) Cloning and characterization of the gene encoding the 28-kDa glutathione S-transferase of Schistosoma mansoni. Gene 24:245–249

    Article  Google Scholar 

  • Reddy CC, Scholz RW, Massaro EJ (1981) Cadmium, methylmercury, mercury, and lead inhibition of calf liver glutathione S-transferase exhibiting selenium-independent glutathione peroxidase activity. Toxicol Appl Pharmacol 61:460–468

    Article  PubMed  CAS  Google Scholar 

  • Rossjohn J, Feil SC, Wilce MCJ, Sexton JL, Spithill TW, Parker MW (1997) Crystallization, structural determination and analysis of a novel parasite vaccine candidate: Fasciola hepatica glutathione S-transferase. J Mol Biol 273:857–872

    Article  PubMed  CAS  Google Scholar 

  • Salvatore L, Wijffels G, Sexton JL, Panaccio M, Mailer S, McCauley I, Spithill TW (1995) Biochemical analysis of recombinant glutathione S-transferase of Fasciola hepatica. Mol Biochem Parasitol 69:281–288

    Article  PubMed  CAS  Google Scholar 

  • Sexton JL, Milner AR, Panaccio M, Waddington J, Wijffels G, Chandler D, Thompson C, Wilson L, Spithill TW, Mitchell GF, Campbell NJ (1990) Glutathione S-transferase: novel vaccine against Fasciola hepatica infection in sheep. J Immunol 145:3905–3911

    PubMed  CAS  Google Scholar 

  • Sheehan D, Meade G, Foley VM, Dowd CA (2001) Structure, function and evolution of glutathione transferases: implications for classification of non-mammalian members of an ancient enzyme superfamily. Biochem J 360:1–16

    Article  PubMed  CAS  Google Scholar 

  • Sluis-Cremer N, Naidoo NN, Kaplan WH, Manoharan TH, Fahl WE, Dirr HW (1996) Determination of a binding site for a non-substrate ligand in mammalian cytosolic glutathione S-transferase by means of fluorescence-resonance energy transfer. Eur J Biochem 241:484–488

    Article  PubMed  CAS  Google Scholar 

  • Smooker PM, Steeper KR, Drew DR, Strugnell RA, Spithill TW (1999) Humoral responses in mice following vaccination with DNA encoding glutathione S-transferase of Fasciola hepatica: effects of mode of vaccination and the cellular compartment of antigen expression. Parasite Immunol 21:357–364

    Article  PubMed  CAS  Google Scholar 

  • Townsend AJ, Goldsmith ME, Pickett CB, Cowan KH (1989) Isolation, characterization, and expression in Escherichia coli of two murine Mu class glutathione S-transferase cDNAs homologous to the rat subunits 3 (Yb1) and 4 (Yb2). J Biol Chem 264:21582–21590

    PubMed  CAS  Google Scholar 

  • Trottein F, Kieny MP, Verwaerde C, Torpier G, Pierce RJ, Balloul JM, Schimitt D, Lecocq JP, Capron A (1990) Molecular cloning and tissue distribution of a 26 kDa Schistosoma mansoni glutathione S-transferase. Mol Biochem Parasitol 41:35–44

    Article  PubMed  CAS  Google Scholar 

  • Walker J, Moreman AD, Crowley P, Barrett J (1993) Biochemical properties of cloned Glutathione S-transferase from Schistosoma mansoni and Schistosoma japonicum. Mol Biochem Parasitol 61:255–264

    Article  PubMed  CAS  Google Scholar 

  • Wijffels GL, Sexton JL, Salvatore L, Pettett JM, Humphris DC, Panaccio M, Spithill TW (1992) Primary sequence heterogeneity and tissue expression of glutathione S-transferases of Fasciola hepatica. Exp Parasitol 74:87–99

    Article  PubMed  CAS  Google Scholar 

  • Xu CB, Verwaerde C, Gras-Masse H, Fontaine J, Bossus M, Trottein F, Wolowczuk I, Tartar A, Capron A (1993) Schistosoma mansoni 28-kDa glutathione S-transferase and immunity against parasite fecundity and egg viability. J Immunol 150:940–949

    PubMed  CAS  Google Scholar 

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Authors and Affiliations

  1. Department of Parasitology, Chung-Ang University College of Medicine, Seoul, 156-756, South Korea

    Ji-Yun Lee, Tae Im Kim, Ki Ho Moon, Shin-Yong Kang & Sung-Jong Hong

  2. Department of Infection Biology, College of Medicine, Chungnam National University, Daejeon, 301-747, South Korea

    Tae Yun Kim

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  1. Tae Yun Kim
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  2. Ji-Yun Lee
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  3. Tae Im Kim
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Correspondence to Sung-Jong Hong.

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Kim, T.Y., Lee, JY., Kim, T.I. et al. Molecular cloning and enzymatic characterization of a class mu glutathione S-transferase of Paragonimus westermani . Parasitol Res 101, 1225–1231 (2007). https://doi.org/10.1007/s00436-007-0626-4

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  • DOI: https://doi.org/10.1007/s00436-007-0626-4

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