Chapters authored
The Role of Glutathione Transferases in the Development of Insecticide Resistance
Glutathione transferases are multifunctional enzymes. Some of the known functions of the enzymes are biotransformation of xenobiotics, countering oxidative stress and participating in cell regulatory functions. As the isoforms present in number of classes the purification of a particular isoform for characterization is a challenging task. In insect, the study of GSTs is focusing on their roles in development of insecticide resistance. There were evident that certain classes of the enzymes are reactive towards conjugating the pesticides. This makes GSTs one of the enzymes of intention in the discipline of pesticide control management.
Part of the book: Insecticides Resistance
Substrate Specificities and Kinetic Parameters of Recombinant Drosophila melanogaster Glutathione S-Transferases E6 and E7
D. melanogaster glutathione transferases E6 and E7 (DmGSTE6 and DmGSTE7) were successfully cloned, purified, and biochemically characterized. The recombinant proteins were readily purified using the combination of both anionic and BSP/GSH-agarose affinity chromatography. Although both GSTs have significant identity in their amino acid sequence, each enzyme displayed unique biochemical characteristics. Both recombinant proteins were only active toward 1-chloro-2,4-dinitrobenzene (CDNB), 1,2-dichloro-4-nitrobenzene (DCNB), and p-nitrobenzyl chloride (p-NBC) with significant difference in catalytic activities. The findings have shown that neither GSTE6 nor GSTE7 was able to counter oxidative stress. Comparatively, GSTE7 was a more efficient enzyme at turning over DCNB and p-NBC, based on its kcat/Km values which were of 0.183 and 2.25 min−1 mM−1, respectively. Thin-layer chromatography analysis showed that both isoforms were not able to conjugate several tested insecticides. The inhibition kinetics of natural products and dyes toward GSTs in vitro revealed that phenol red possessed inhibition effects only on GSTE6 while rose bengal and cardiogreen inhibit significantly on both GSTE6 and GSTE7. In contrast, methylene blue dye and trans-chalcone have been shown to stimulate GSTE7 activity toward CDNB.
Part of the book: Drosophila melanogaster