Hostname: page-component-cd9895bd7-hc48f Total loading time: 0 Render date: 2024-12-28T21:49:10.752Z Has data issue: false hasContentIssue false
  • English
  • Français

Molecular cloning and characterization of the copper/zinc and manganese superoxide dismutase genes from the human parasite Clonorchis sinensis

Published online by Cambridge University Press:  24 January 2005

A. H. LI ,
Y. KONG ,
S. H. CHO ,
H. W. LEE ,
B. K. NA ,
J. K. PAK  and
T. S. KIM
A. H. LI
Affiliation:
Department of Tropical and Endemic Parasitic Diseases, National Institute of Health, Seoul 122-701, Korea State Key Laboratory of Freshwater Ecology and Biotechnology, Institute of Hydrobiology, Chinese Academy of Sciences, Wuhan, 430072, China
Y. KONG
Affiliation:
Department of Molecular Parasitology, Sungkyunkwan University School of Medicine, Suwon 440-746, Korea
S. H. CHO
Affiliation:
Department of Tropical and Endemic Parasitic Diseases, National Institute of Health, Seoul 122-701, Korea
H. W. LEE
Affiliation:
Department of Tropical and Endemic Parasitic Diseases, National Institute of Health, Seoul 122-701, Korea
B. K. NA
Affiliation:
Department of Molecular Parasitology, Sungkyunkwan University School of Medicine, Suwon 440-746, Korea
J. K. PAK
Affiliation:
Asan Institute for Life Sciences, College of Medicine, University of Ulsan, Seoul 138-736, Korea
T. S. KIM
Affiliation:
Department of Tropical and Endemic Parasitic Diseases, National Institute of Health, Seoul 122-701, Korea
Get access Rights & Permissions [Opens in a new window]

Abstract

A copper/zinc superoxide dismutase (Cu/ZnSOD) gene and a manganese superoxide dismutase (MnSOD) gene of the human parasite Clonorchis sinensis have been cloned and their gene products functionally characterized. Genes Cu/ZnSOD and MnSOD encode proteins of 16 kDa and 25·4 kDa, respectively. The deduced amino acid sequences of the two genes contained highly conserved residues required for activity and secondary structure formation of Cu/ZnSOD and MnSOD, respectively, and show up to 73·7% and 75·4% identities with their counterparts in other animals. The genomic DNA sequence analysis of Cu/ZnSOD gene revealed this as an intronless gene. Inhibitor studies with purified recombinant Cu/ZnSOD and MnSOD, both of which were functionally expressed in Escherichia coli, confirmed that they are copper/zinc and manganese-containing SOD, respectively. Immunoblots showed that both C. sinensis Cu/ZnSOD and MnSOD should be antigenic for humans, and both, especially the C. sinensis MnSOD, exhibit extensive cross-reactions with sera of patients infected by other trematodes or cestodes. RT-PCR and SOD activity staining of parasite lysates indicate that there are no significant differences in mRNA level or SOD activity for both species of SOD, indicating cytosolic Cu/ZnSOD and MnSOD might play a comparatively important role in the C. sinensis antioxidant system.

Keywords

Clonorchis sinensiscopper/zinc superoxide dismutasemanganese superoxide dismutasecDNAgenomic DNArecombinant enzymehelminths
Type
Research Article
Information
Parasitology , Volume 130 , Issue 6 , June 2005 , pp. 687 - 697
Copyright
© 2005 Cambridge University Press

Access options

Get access to the full version of this content by using one of the access options below. (Log in options will check for institutional or personal access. Content may require purchase if you do not have access.)

References

REFERENCES

BEAUCHAMP, C. & FRIDOVICH, I. ( 1971). Superoxide dismutase improved assays and assay applicable to acrylamide gels. Analytical Biochemistry 44, 276–287.CrossRefGoogle Scholar
BRADFORD, M. M. ( 1976). A rapid and sensitive method for the quantification of microgram quantities of protein utilizing the principle of protein-dye binding. Analytical Biochemistry 72, 248–250.CrossRefGoogle Scholar
BRITTON, C., KNOX, D. P. & KENNEDY, M. W. ( 1994). Superoxide dismutase (SOD) activity of Dictyocaulus viviparus and its inhibition by antibody from infected and vaccinated bovine hosts. Parasitology 109, 257–263.CrossRefGoogle Scholar
BUTTERWORTH, A. E. ( 1984). Cell-mediated damage to helminths. Advances in Parasitology 23, 143–235.Google Scholar
CALLAHAN, H. L., CROUCH, R. K. & JAMES, E. R. ( 1991). Dirofilaria immitis superoxide dismutase: purification and characterization. Molecular and Biochemical Parasitology 49, 245–252.CrossRefGoogle Scholar
CARVALHO-QUEIROZ, C., COOK, R., WANG, C. C., CORREA-OLIVEIRA, R., BAILEY, N. A., EGILMEZ, N. K., MATHIOWITZ, E. & LOVERDE, P. T. ( 2004). Cross-reactivity of Schistosoma mansoni cytosolic superoxide dismutase, a protective vaccine candidate, with host superoxide dismutase and identification of parasite-specific B epitopes. Infection and Immunity 72, 2635–2647.CrossRefGoogle Scholar
CHEN, J. R., WENG, C. N., HO, T. Y., CHENG, I. C. & LAI, S. S. ( 2000). Identification of the copper-zinc superoxide dismutase activity in Mycoplasma hyopneumoniae. Veterinary Microbiology 73, 301–310.CrossRefGoogle Scholar
CHOI, D. H., NA, B. K., SEO, M. S., SONG, H. R. & SONG, C. Y. ( 2000). Purification and characterization of iron superoxide dismutase and copper-zinc superoxide dismutase from Acanthamoeba castellanii. Journal of Parasitology 86, 899–907.CrossRefGoogle Scholar
CHUNG, Y. B., SONG, C. Y., LEE, H. S., KONG, Y. & CHO, S. Y. ( 1991). Purification and characterization of a Cu, Zn-superoxide dismutase from adult Paragonimus westermani. Korean Journal of Parasitology 29, 259–266.CrossRefGoogle Scholar
CIRIOLO, M. R., BATTISTONI, A., FALCONI, M., FILOMENI, G. & ROTILIO, G. ( 2001). Role of the electrostatic loop of Cu,Zn superoxide dismutase in the copper uptake process. European Journal of Biochemistry 268, 737–742.CrossRefGoogle Scholar
CROMPTON, D. W. ( 1999). How much human helminthiasis is there in the world? Journal of Parasitology 85, 397–403.Google Scholar
FRIDOVICH, I. ( 1978). The biology of oxygen radicals. Science 201, 875–880.CrossRefGoogle Scholar
GELLER, B. L. & WINGE, D. R. ( 1983). A method for distinguishing Cu,Zn- and Mn-containing superoxide dismutases. Analytical Biochemistry 128, 86–92.CrossRefGoogle Scholar
GOLENSER, J., PELED-KAMAR, M., SCHWARTZ, E., FRIEDMAN, I., GRONER, Y. & POLLACK, Y. ( 1998). Transgenic mice with elevated level of CuZnSOD are highly susceptible to malaria infection. Free Radical Biology and Medicine 24, 1504–1510.CrossRefGoogle Scholar
HENKLE, K. J., LIEBAU, E., MÜLLER, S., BERGMANN, B. & WALTER, R. D. ( 1991). Characterization and molecular cloning of a Cu/Zn superoxide dismutase from the human parasite Onchocerca volvulus. Infection and Immunity 59, 2063–2069.Google Scholar
HENKLE-DÜHRSEN, K., TAWE, W., WARNECKE, C. & WALTER, R. D. ( 1995). Characterization of the manganese superoxide dismutase cDNA and gene from the human parasite Onchocerca volvulus. The Biochemical Journal 308, 441–446.CrossRefGoogle Scholar
HESS, J., DIETRICH, G., GENTSCHEV, I., MIKO, D., GOEBEL, W. & KAUFMANN, S. H. ( 1997). Protection against murine listeriosis by an attenuated recombinant Salmonella typhimurium vaccine strain that secretes the naturally somatic antigen superoxide dismutase. Infection and Immunity 65, 1286–1292.Google Scholar
HONG, Z., KOSMAN, D. J., THAKUR, A., REKOSH, D. & LOVERDE, P. T. ( 1992). Identification and purification of a second form of Cu/Zn superoxide dismutase from Schistosoma mansoni. Infection and Immunity 60, 3641–3651.Google Scholar
HONG, Z., LOVERDE, P. T., HAMMARSKJOLD, M. L. & REKOSH, D. ( 1992). Schistosoma mansoni: cloning of a complementary DNA encoding a cytosolic Cu/Zn superoxide dismutase and high-yield expression of the enzymatically active gene product in Escherichia coli. Experimental Parasitology 75, 308–322.CrossRefGoogle Scholar
HONG, Z., LOVERDE, P. T., THAKUR, A., HAMMARSKJOLD, M. L. & REKOSH, D. ( 1993). Schistosoma mansoni: a Cu/Zn superoxide dismutase is glycosylated when expressed in mammalian cells and localized to a subtegumental region in adult schistosomes. Experimental Parasitology 76, 101–114.CrossRefGoogle Scholar
HUNTER, T., BANNISTER, W. H. & HUNTER, G. J. ( 1997). Cloning, expression and characterization of two manganese superoxide dismutases from Caenorhabditis elegans. Journal of Biological Chemistry 272, 28652–28659.CrossRefGoogle Scholar
IMLAY, J. A. & LINN, S. ( 1988). DNA damage and oxygen radical toxicity. Science 240, 1302–1308.CrossRefGoogle Scholar
IMLAY, K. R. & IMLAY, J. A. ( 1996). Cloning and analysis of sodC, encoding the copper-zinc superoxide dismutase of Escherichia coli. Journal of Bacteriology 178, 2564–2571.CrossRefGoogle Scholar
ISMAIL, S. O., PARAMCHUK, W., SKEIKY, Y. A., REED, S. G., BHATIAM, A. & GEDAMU, L. ( 1997). Molecular cloning and characterization of two iron superoxide dismutase cDNAs from Trypanosoma cruzi. Molecular and Biochemical Parasitology 86, 187–197.CrossRefGoogle Scholar
JAMES, E. R., McLEAN, D. C. Jr & PERLER, F. ( 1994). Molecular cloning of an Onchocerca volvulus extracellular Cu-Zn superoxide dismutase. Infection and Immunity 62, 713–716.Google Scholar
KIM, T. S., JUNG, Y. H., NA, B. K., KIM, K. S. & CHUNG, P. R. ( 2000). Molecular cloning and expression of Cu/Zn-containing superoxide dismutase from Fasciola hepatica. Infection and Immunity 68, 3941–3948.CrossRefGoogle Scholar
LAEMMLI, U. K. ( 1970). Cleavage of structural proteins during assembly of the head of bacteriophage T4. Nature, London 227, 680–685.CrossRefGoogle Scholar
MAIZELS, R. A., BUNDY, D. A. P., SELKIRK, M. E., SMITH, D. F. & ANDERSON, R. M. ( 1993). Immunological modulation and evasion by helminth parasites in human populations. Nature, London 365, 797–805.CrossRefGoogle Scholar
MARRES, C. A., VAN LOON, A. P., OUDSHOORN, P., VAN STEEG, H., GRIVELL, L. A. & SLATER, E. C. ( 1985). Nucleotide sequence analysis of the nuclear gene coding for manganese superoxide dismutase of yeast mitochondria, a gene previously assumed to code for the Rieske iron-sulphur protein. European Journal of Biochemistry 147, 153–161.CrossRefGoogle Scholar
MURRAY, H. W. ( 1981). Susceptibility of Leishmania to oxygen intermediates and killing by normal macrophages. Journal of Experimental Medicine 153, 1302–1315.CrossRefGoogle Scholar
NIELSEN, H., ENGELBRECHT, J., BRUNAK, S. & VON HEIJNE, G. ( 1997). Identification of prokaryotic and eukaryotic signal peptides and prediction of their cleavage sites. Protein Engineering 10, 1–6.CrossRefGoogle Scholar
OU, X., TANG, L., McCROSSAN, M., HENKLE-DUHRSEN, K. & SELKIRK, M. E. ( 1995). Brugia malayi: localization and differential expression of extracellular and cytoplasmic Cu, Zn superoxide dismutases in adults and microfilariae. Experimental Parasitology 80, 515–529.CrossRefGoogle Scholar
PARAMCHUK, W. J., ISMAIL, S. O., BHATIA, A. & GEDAMU, L. ( 1997). Cloning, characterization and overexpression of two iron superoxide dismutase cDNAs from Leishmania chagasi: role in pathogenesis. Molecular and Biochemical Parasitology 90, 203–221.CrossRefGoogle Scholar
RIM, H. J. ( 1986). Current pathobiology and chemotherapy of clonorchiasis. Korean Journal of Parasitology 24 (Suppl.), S1–S141.CrossRefGoogle Scholar
TANG, L., OU, X., HENKLE-DURHRSEN, K. & SELKIRK, M. E. ( 1994). Extracellular and cytoplasmic Cu/Zn superoxide dismutase from Brugia lymphatic filarial nematode parasites. Infection and Immunity 62, 961–967.Google Scholar