Kang, TH Yun, DH Lee, EHB Chung, YB Bae, YA Chung, JY Kang, I Kim, J Cho, SY Kong, Y 2024-01-21T07:38:21Z 2024-01-21T07:38:21Z 2021-09-02 2004-02 0031-1820 https://pubs.kist.re.kr/handle/201004/137909 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 E 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. English CAMBRIDGE UNIV PRESS CYSTEINE PROTEASE INHIBITORS FASCIOLA-HEPATICA MOLECULAR-CLONING PROTEINASE EXPRESSION GENE PAPAIN FAMILY DOMAIN PURIFICATION A cathepsin F of adult Clonorchis sinensis and its phylogenetic conservation in trematodes Article 10.1017/S0031182003004335 1 PARASITOLOGY, v.128, pp.195 - 207 PARASITOLOGY 128 195 207 scie scopus 000220513500010 2-s2.0-10744229736 Parasitology Parasitology Article CYSTEINE PROTEASE INHIBITORS FASCIOLA-HEPATICA MOLECULAR-CLONING PROTEINASE EXPRESSION GENE PAPAIN FAMILY DOMAIN PURIFICATION Clonorchis sinensis excretory-secretory cysteine protease cathepsin F ERFNAQ motif E/DXGTA motif recombinant antigen