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

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.

Life span and stress resistance of Caenorhabditis elegans are differentially affected by glutathione transferases metabolizing 4-hydroxynon-2-enal

“…11) -are separated by considerable evolutionary distance. This situation resembles that found by us in the fruitfly Drosophila melanogaster where 4-HNE-metabolizing GSTs belong to at least two distinct classes Sawicki et al, 2003). The pattern depicted in Fig.…”

Antimicrobial Peptides Increase Tolerance to Oxidant Stress in Drosophila melanogaster

“…Consistent with this scenario, we observed decreased JNK signaling and increased GST activity in the flies with Dpt overexpression and confirmed these changes by real-time PCR and enzyme activity assays, respectively. GST has been shown to play a role in antioxidant defense and has glutathione reductase activity in Drosophila (20,22,54). In particular, GstD3, which was up-regulated in the flies with Dpt overexpression, has also been shown to respond to dietary H 2 O 2 , and both GstD3 and GstE1 can metabolize 4-hydroxynonenal, a highly toxic aldehyde produced by lipid peroxidation in cells in response to oxidative stress (54,55).…”

Identification, genomic organization, and oxidative stress response of a sigma class glutathione S-transferase gene (AccGSTS1) in the honey bee, Apis cerana cerana

“…Earlier studies have demonstrated that paraquat treatment results in oxidative stress and induces the expression of several classes of GST genes in C. riparius . Similarly, cumene hydroperoxide and t-butylhydroperoxide are also widely used as model substances for studying the mechanism of cell injury resulting from oxidative stress (Sawicki et al 2003;Tang and Tu 1994). In the present study, paraquat, t-butylhydroperoxide and cumene hydroperoxide adsorbed to the filter disc may have induced oxidative stress in E. coli cells.…”

“…The N-terminal domain (βαβαββα motif) containing the G-site and the C-terminal domain (all α-helical motifs) harboring the H-site are shown. The conserved Tyr at position 8 responsible for the stabilization of GSH is indicated oxidative stress (Arrese and Soulages 2010;Enayati et al 2005;Sawicki et al 2003). In addition, there is evidence that insect GSTs can detoxify many plant allelochemicals and synthetic insecticides (Li et al 2007).…”

“…Because GST genes play important roles in detoxification pathways and the oxidative stress response, studies on GST resistance have been widely reported in insects, including Drosophila melanogaster Low et al 2007;Sawicki et al 2003;Tang and Tu 1994;Tu and Akgul 2005), Anopheles gambiae (Chen et al 2003;Ding et al 2003Ding et al , 2005Ranson et al 2000Ranson et al , 2001, and Bombyx mori (Yamamoto et al 2006;Yu et al 2008), but information about other important species is rather limited. Honey bees, which serve as an excellent model for social behavior research, are a major plant pollinator and thus play essential roles in agriculture and ecological balance.…”