Molecular cloning, expression, biochemical characteristics, and biomarker potential of theta class glutathione S-transferase (GST-T) from the polychaete Neanthes succinea

Aquat Toxicol. 2007 Jun 15;83(2):104-15. doi: 10.1016/j.aquatox.2007.03.015. Epub 2007 Mar 27.

Abstract

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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Animals
  • Base Sequence
  • Biomarkers / analysis
  • Cloning, Molecular
  • Copper / toxicity
  • DNA, Complementary / chemistry
  • Dinitrochlorobenzene / metabolism
  • Ethylmaleimide / pharmacology
  • Gene Expression Regulation, Enzymologic* / drug effects
  • Geologic Sediments / analysis
  • Glutathione / metabolism
  • Glutathione Transferase / analysis
  • Glutathione Transferase / biosynthesis*
  • Glutathione Transferase / genetics*
  • Glutathione Transferase / metabolism
  • Hemin / pharmacology
  • Hydrogen-Ion Concentration
  • Molecular Sequence Data
  • Phylogeny
  • Polychaeta / classification
  • Polychaeta / enzymology*
  • Polychaeta / genetics*
  • Recombinant Proteins / analysis
  • Recombinant Proteins / biosynthesis
  • Temperature
  • Triazines / pharmacology
  • Water Pollutants, Chemical / toxicity

Substances

  • Biomarkers
  • DNA, Complementary
  • Dinitrochlorobenzene
  • Recombinant Proteins
  • Triazines
  • Water Pollutants, Chemical
  • Cibacron Blue F 3GA
  • Hemin
  • Copper
  • cuprous chloride
  • Glutathione Transferase
  • Glutathione
  • Ethylmaleimide

Associated data

  • GENBANK/EF394324