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.
Wu
Zhongluan
Z
Department of Parasitology, School of Pre-Clinical Medicine, SUN Yat-Sen University, Guangzhou, 510089, China.
Wu
De
D
Hu
Xuchu
X
Xu
Jin
J
Chen
Shouyi
S
Wu
Zhongdao
Z
Yu
Xinbing
X
D015766:Albendazole / Q000494:pharmacology; D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D003001:Cloning, Molecular; D003004:Clonorchis sinensis / Q000201:enzymology* / Q000235:genetics*; D018076:DNA, Complementary; D018509:DNA, Helminth / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification; D004137:Dinitrochlorobenzene / Q000378:metabolism; D004791:Enzyme Inhibitors / Q000494:pharmacology; D004926:Escherichia coli; D005982:Glutathione Transferase / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism*; D008969:Molecular Sequence Data; D016366:Open Reading Frames; D011994:Recombinant Proteins / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D017422:Sequence Analysis, DNA; D017386:Sequence Homology, Amino Acid; D014227:Triazines / Q000494:pharmacology
D018076:DNA, Complementary; D018509:DNA, Helminth; D004137:Dinitrochlorobenzene; D004791:Enzyme Inhibitors; D011994:Recombinant Proteins; D014227:Triazines; C012842:Cibacron Blue F 3GA; D005982:Glutathione Transferase; D015766:Albendazole