Cloning, expression and biochemical characterization of one epsilon-class (GST-3) and ten Delta-class (GST-1) glutathione S-transferases from Drosophila melanogaster, and identification of additional nine members of the epsilon class

Rafał Sawicki, Sharda P. Singh, Ashis K. Mondal, Helen Beneš, Piotr Zimniak

Research output: Contribution to journalArticlepeer-review

178 Scopus citations

Abstract

From the fruitfly, Drosophila melanogaster, ten members of the cluster of Delta-class glutathione S-transferases (GSTs; formerly denoted as Class I GSTs) and one member of the Epsilon-class cluster (formerly GST-3) have been cloned, expressed in Escherichia coli, and their catalytic properties have been determined. In addition, nine more members of the Epsilon cluster have been identified through bioinformatic analysis but not further characterized. Of the 11 expressed enzymes, seven accepted the lipid peroxidation product 4-hydroxynonenal as substrate, and nine were active in glutathione conjugation of 1-chloro-2,4-dinitrobenzene. Since the enzymically active proteins included the gene products of DmGSTD3 and DmGSTD7 which were previously deemed to be pseudogenes, we investigated them further and determined that both genes are transcribed in Drosophila. Thus our present results indicate that DmGSTD3 and DmGSTD7 are probably functional genes. The existence and multiplicity of insect GSTs capable of conjugating 4-hydroxynonenal, in some cases with catalytic efficiencies approaching those of mammalian GSTs highly specialized for this function, indicates that metabolism of products of lipid peroxidation is a highly conserved biochemical pathway with probable detoxification as well as regulatory functions.

Original languageEnglish (US)
Pages (from-to)661-669
Number of pages9
JournalBiochemical Journal
Volume370
Issue number2
DOIs
StatePublished - Mar 1 2003
Externally publishedYes

Keywords

  • Detoxification
  • Diptera
  • Electrophile
  • Glutathione conjugation
  • Lipid peroxidation

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology
  • Cell Biology

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