Molecular cloning and characterization of cDNA encoding a novel cytosolic glutathione transferase from Clonorchis sinensis.

Glutathione transferases (GSTs) are a group of multifunction isoenzymes coded by many genes. A cDNA encoding a novel cytosolic GST enzyme was cloned from a Clonorchis sinensis (Cs) adult worm cDNA library by large-scale sequencing. This new cDNA contains 786 bp with a putative open reading frame of 212 amino acids. The deduced amino acid sequence exhibits 61% identity to C. sinensis cytosolic 28-kDa GST. Recombinant CsGST was overexpressed in Escherichia coli BL21(DE3) and was purified by Ni-NTA Agarose. Enzymatic assays showed that the recombinant CsGST had a high activity of 22.7 U mg(-1). The average K (m) of the CsGST for 1-chloro-2,4-dinitrobenzene is 111 microM. Cibacron blue F3GA and albendazole inhibited the CsGST activity with respective average IC(50) of 1.1 and 247.1 microM. It provides a model for the structure and physiological function analysis on CsGST. The nucleotide sequence reported in this paper has been submitted to the GenBank Database with accession number DQ179264.