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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
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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

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