Molecular cloning, expression, biochemical characteristics, and biomarker potential of theta class glutathione S-transferase (GST-T) from the polychaete Neanthes succinea.
We cloned and sequenced the full-length cDNA of a theta class glutathione S-transferase (GST-T) from the polychaete Neanthes succinea. The open reading frame of N. succinea GST-T cDNA was 678bp and encoded 226 amino acid residues. We generated recombinant N. succinea GST-T by expression in transformed Escherichia coli and studied the kinetic properties as well as the effects of inhibitors, pH, and temperature on N. succinea GST-T. GST-T expression was studied using real-time RT-PCR in response to exposure to the model oxidative stress-inducing agent, CuCl(2). Copper induced a concentration-dependant increase in the expression of GST-T. Moreover, polychaetes collected from a heavily contaminated lake near an industrial complex showed significantly higher levels of GST-T expression. Interestingly, the site-collected polychaetes with the highest GST-T mRNA expression levels also showed the highest metallothioneins levels. These results suggest that GST-T in polychaetes may have an antioxidant role and that N. succinea GST-T expression may be a useful biomarker for exposure to environmental contaminants such as copper. Our findings provide a better understanding of the biochemical characteristics of N. succinea GST-T, and elucidate the potential role of GST-T in heavy metal-induced oxidative stress and as a biomarker for environmental contamination.
Rhee
Jae-Sung
JS
Department of Molecular and Environmental Bioscience, Graduate School, Hanyang University, Seoul, South Korea.
Lee
Young-Mi
YM
Hwang
Dae-Sik
DS
Won
Eun-Ji
EJ
Raisuddin
Sheikh
S
Shin
Kyung-Hoon
KH
Lee
Jae-Seong
JS
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D015415:Biomarkers / Q000032:analysis; D003001:Cloning, Molecular; D003300:Copper / Q000633:toxicity; D018076:DNA, Complementary / Q000737:chemistry; D004137:Dinitrochlorobenzene / Q000378:metabolism; D005033:Ethylmaleimide / Q000494:pharmacology; D015971:Gene Expression Regulation, Enzymologic / Q000187:drug effects; D019015:Geologic Sediments / Q000032:analysis; D005978:Glutathione / Q000378:metabolism; D005982:Glutathione Transferase / Q000032:analysis / Q000096:biosynthesis* / Q000235:genetics* / Q000378:metabolism; D006427:Hemin / Q000494:pharmacology; D006863:Hydrogen-Ion Concentration; D008969:Molecular Sequence Data; D010802:Phylogeny; D011077:Polychaeta / Q000145:classification / Q000201:enzymology* / Q000235:genetics*; D011994:Recombinant Proteins / Q000032:analysis / Q000096:biosynthesis; D013696:Temperature; D014227:Triazines / Q000494:pharmacology; D014874:Water Pollutants, Chemical / Q000633:toxicity
D015415:Biomarkers; D018076:DNA, Complementary; D004137:Dinitrochlorobenzene; D011994:Recombinant Proteins; D014227:Triazines; D014874:Water Pollutants, Chemical; C012842:Cibacron Blue F 3GA; D006427:Hemin; D003300:Copper; C028419:cuprous chloride; D005982:Glutathione Transferase; D005978:Glutathione; D005033:Ethylmaleimide