title: Purification and biochemical characterization of glutathione S-transferases from four field populations of Bactrocera dorsalis (Hendel) (Diptera: Tephritidae).
abstract: Glutathione S-transferases (GSTs) are a group of detoxification enzymes that catalyze the nucleophilic addition of glutathione to a wide variety of endogenous and xenobiotic compounds. In this study, GSTs were purified from four field populations of Bactrocera dorsalis with different insecticide susceptibilities by glutathione-agarose affinity chromatography. The populations were collected from Dongguan (DG) and Guangzhou (GZ) of the Guangdong Province, Haikou of the Hainan province (HN), and Kunming of the Yunnan province (YN), China. Differences in GST characteristics among the four populations were studied using purified enzyme samples through comparative SDS-PAGE, kinetic, and inhibition experiments. The specific activities of the purified enzymes were similar, but the purification yield of the GZ population (31.54%) was the lowest. SDS-PAGE analysis showed only one band at approximately 23 kDa for these four populations. Kinetic analyses showed that the affinities of the purified GSTs from the GZ and YN populations for 1-chloro-2.4-dinitrobenzene (CDNB) were much higher than those of GSTs from the other two populations, whereas the HN population had the highest catalytic capability in terms of V(max) value. The optimum temperature for CDNB conjugation was 37 °C and the optimum pH was 7.5 in all four populations. Inhibition kinetics showed that ethacrynic acid, diethyl maleate, tetraethylthiuram disulfide, curcumin, bromosulfalein, and β-cypermethrin had excellent inhibitory effects on GSTs in the four populations of B. dorsalis, but the low inhibitory effects of malathion and avermectin did not differ between populations. These results suggest that GSTs may have a role in detoxification of β-cypermethrin in B. dorsalis.
authors: Key Laboratory of Entomology and Pest Control Engineering, College of Plant Protection, Southwest University, Chongqing 400715, People's Republic of China.
authors: Fei
authors: F
authors: Hu
authors: Wei
authors: W
authors: Dou
authors: Jing-Jing
authors: JJ
authors: Wang
authors: Fu-Xian
authors: FX
authors: Jia
authors: Jin-Jun
authors: JJ
authors: Wang
mesh_terms: D000818:Animals; D005982:Glutathione Transferase / Q000737:chemistry / Q000302:isolation & purification*; D008658:Inactivation, Metabolic; D019476:Insect Proteins / Q000737:chemistry / Q000302:isolation & purification*; D007305:Insecticide Resistance; D007306:Insecticides / Q000493:pharmacokinetics*; D007700:Kinetics; D033621:Tephritidae / Q000201:enzymology*
chemical_list: D019476:Insect Proteins; D007306:Insecticides; D005982:Glutathione Transferase