Purification and partial characterization of glutathione S-transferase from insecticide-resistant field populations of Liposcelis paeta Pearman (Psocoptera: Liposcelididae).
Enzymes that possess glutathione S-transferase (GST) activity were purified to homogeneity by glutathione-agarose affinity chromatography from three field populations of Liposcelis paeta (Pearman). These populations were collected from Nanyang city of Henan Province (NY), Wuzhou (WZ) and Hezhou (HZ) cities of Guangxi Province, China, and had different susceptibilities to dichlorvos [LC(50)s of the NY (281.48 mg/m(2)), the WZ (285.07 mg/m(2)), and the HZ (243.52 mg/m(2)), respectively]. The specific activities of purified enzymes from these three populations increased 32.24-, 99.81-, and 42.52-fold, respectively. Kinetic analyses showed that the catalytic activity of purified GST from NY population towards GSH was much higher than the others, while WZ population reached the highest in V(max) (CDNB). SDS-polyacrylamide electrophoresis revealed that the purified GST had two subunits with a molecular mass of 23.31 and 20.43 kDa for NY, 53.14 and 20.13 kDa for WZ, and 50.79 and 19.42 kDa for HZ, respectively. The in vitro inhibition studies of GSTs indicated that three kinds of insecticides (chlorpyrifos, carbosulfan, and cypermethrin) and five metallic ions (Zn(2+), Ba(2+), Ca(2+), Hg(2+), Mn(2+), and Mg(2+)) all possessed inhibitory effects on purified GST, and ethacrynic acid (EA, a specific inhibitor of GST) expressed inhibitory effects. In the bioassay, three populations of L. paeta had different susceptibilities to different insecticides, even after they were reared on diets consisting of 25% EA. The GST activities of L. paeta from different areas also showed different temperature and pH stabilities. The differences in GST among the three populations may be attributed partially to the differences in control practices for psocids between Henan and Guangxi Provinces. (c) 2009 Wiley Periodicals, Inc.
Wu
Shuang
S
Key Laboratory of Entomology and Pest Control Engineering, College of Plant Protection, Southwest University, Chongqing 400715, P. R. China.
Dou
Wei
W
Wu
Jing-Jing
JJ
Wang
Jin-Jun
JJ
D000818:Animals; D002412:Cations / Q000378:metabolism; D004591:Electrophoresis, Polyacrylamide Gel; D005982:Glutathione Transferase / Q000037:antagonists & inhibitors / Q000302:isolation & purification* / Q000378:metabolism; D006863:Hydrogen-Ion Concentration; D007313:Insecta / Q000201:enzymology*; D007305:Insecticide Resistance; D007700:Kinetics; D013696:Temperature; D018675:Toxicity Tests
D002412:Cations; D005982:Glutathione Transferase