Stimulatory effect of insecticides on partially purified P-glycoprotein ATPase from the resistant pest Helicoverpa armigera.
A P-glycoprotein-like protein (Ha-Pgp) was detected in a membrane preparation from the insecticide-resistant pest Helicoverpa armigera (Lepidoptera: Noctüidae) using C219 antibodies that are directed towards an epitope in the nucleotide-binding domains. This protein was partially purified and found to be a glycoprotein displaying ATPase activity. SDS-PAGE confirmed that a high molecular mass glycoprotein (150 kDa) was overexpressed in resistant pests, but was not detected in susceptible pests. The partially purified Ha-Pgp ATPase was reconstituted into proteoliposomes and it was found that some insecticides, namely, monocrotophos, endosulfan, cypermethrin, fenvalerate, and methylparathion, stimulated the ATPase activity. The effect of various inhibitors on partially purified Ha-Pgp showed that orthovanadate is a potent inhibitor of its ATPase activity, inhibiting it by 90% at a concentration of 2 mmol/L. Other inhibitors, such as EDTA, sodium azide, and molybdate resulted in only a 20% decrease in activity. Details of the structure and function of Ha-Pgp will be important in the development of strategies to overcome insecticide resistance in this pest.
Aurade
Ravindra
R
Department of Biochemistry, Gulbarga University, Gulbarga 585 106, India.
Jayalakshmi
Senigala K
SK
Sreeramulu
Kuruba
K
D018435:ATP Binding Cassette Transporter, Subfamily B / Q000378:metabolism*; D000251:Adenosine Triphosphatases / Q000302:isolation & purification* / Q000378:metabolism*; D000818:Animals; D007305:Insecticide Resistance / Q000235:genetics / Q000502:physiology; D007306:Insecticides / Q000494:pharmacology*; D009036:Moths / Q000187:drug effects* / Q000201:enzymology*
D018435:ATP Binding Cassette Transporter, Subfamily B; D007306:Insecticides; D000251:Adenosine Triphosphatases