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A cathepsin F of adult Clonorchis sinensis and its phylogenetic conservation in trematodes

Published online by Cambridge University Press:  01 March 2004

T. H. KANG ,
D.-H. YUN ,
E.-H. B. LEE ,
Y.-B. CHUNG ,
Y.-A. BAE ,
J.-Y. CHUNG ,
I. KANG ,
J. KIM ,
S.-Y. CHO  and
Y. KONG
T. H. KANG
Affiliation:
Department of Biotechnology, Korea University Graduate School, Seoul 136-701, Korea Biomedical Research Center, Korea Institute of Science and Technology, Seoul 136-791, Korea
D.-H. YUN
Affiliation:
Department of Molecular Parasitology and Center for Molecular Medicine, Sungkyunkwan University School of Medicine and Samsung Biomedical Research Institute, Suwon 440-746, Korea
E.-H. B. LEE
Affiliation:
Biomedical Research Center, Korea Institute of Science and Technology, Seoul 136-791, Korea
Y.-B. CHUNG
Affiliation:
Department of Parasitology, College of Medicine, Cheju National University, Jeju 690-756, Korea
Y.-A. BAE
Affiliation:
Department of Molecular Parasitology and Center for Molecular Medicine, Sungkyunkwan University School of Medicine and Samsung Biomedical Research Institute, Suwon 440-746, Korea
J.-Y. CHUNG
Affiliation:
Department of Molecular Parasitology and Center for Molecular Medicine, Sungkyunkwan University School of Medicine and Samsung Biomedical Research Institute, Suwon 440-746, Korea
I. KANG
Affiliation:
Department of Biochemistry, Kyunghee University College of Medicine, Seoul 130-701, Korea
J. KIM
Affiliation:
Department of Biotechnology, Korea University Graduate School, Seoul 136-701, Korea
S.-Y. CHO
Affiliation:
Department of Molecular Parasitology and Center for Molecular Medicine, Sungkyunkwan University School of Medicine and Samsung Biomedical Research Institute, Suwon 440-746, Korea
Y. KONG
Affiliation:
Department of Molecular Parasitology and Center for Molecular Medicine, Sungkyunkwan University School of Medicine and Samsung Biomedical Research Institute, Suwon 440-746, Korea
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Abstract

A novel 28 kDa cysteine protease (Cs28CF) secreted by the hepatobiliary trematode, Clonorchis sinensis was identified. The protease was purified from the excretory-secretory products (ESP) of the adult worm using DEAE-ion exchange and Arginine-Sepharose 4B chromatography. It showed a high activity between pH 6·5 and 7·5 in a dithiothreitol (DTT)-dependent manner. Inhibitors specific to cysteine proteases down-regulated the activity. Addition of Cs28CF to monkey cholangiocyte cultures resulted in approximately 95% cell death after 7 days. The full-length cDNA (1078 bp) encoded a single peptide of 328 amino acids (aa) with an N-terminal hydrophobic sequence, an ERFNAQ motif in the propeptide and a mature domain. Expression of mRNA transcripts of Cs28CF was observed in both the metacercaria and adult stages. Bacterially expressed recombinant protein exhibited a specific antibody reaction with clonorchiasis sera. Deduced aa exhibited 52–76% sequence identity with the cathepsin F analogues from other organisms. A novel E/DXGTA motif was recognized in the propeptide region. Phylogenetic analysis of 63 papain family members revealed that the trematode cysteine proteases formed 2 major clades of cathepsins F and L. The trematode cysteine proteases classified as cathepsin F shared higher homology among themselves than those classified as cathepsin L. Cathepsin F is phylogenetically conserved in the trematode parasites as well as in mammals.

Keywords

Clonorchis sinensisexcretory-secretory cysteine proteasecathepsin FERFNAQ motifE/DXGTA motifrecombinant antigen
Type
Research Article
Information
Parasitology , Volume 128 , Issue 2 , February 2004 , pp. 195 - 207
Copyright
2004 Cambridge University Press

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