MODULATION OF P‐GLYCOPROTEIN ATPASE OF HELICOVERPA ARMIGERA BY CHOLESTEROL: EFFECTS ON ATPASE ACTIVITY AND INTERACTION OF INSECTICIDES

Purified P‐glycoprotein ATPase from Helicoverpa armigera (Ha‐Pgp), reconstituted in proteoliposomes composed of phospholipids and cholesterol, shows higher ATPase activity in the presence of cholesterol than in its absence. The Ha‐Pgp ATPase activity was increased 30–40% with cholesterol. The K_M for ATP was found to be 1 and 0.8 mM in the absence and presence of cholesterol, respectively. The insecticide‐stimulated Ha‐Pgp ATPase activity was increased by 10–20% for all the insecticides in the reconstituted proteoliposomes containing cholesterol compared to those with no cholesterol. The effects of cholesterol on K_M and V_max values of insecticide‐stimulated Ha‐Pgp ATPase activity were unrelated to the size of the insecticide. Ha‐Pgp tryptophan fluorescence displayed a red shift of 3 and 8 nm in emission spectra upon binding of insecticides. Cholesterol enhances the interaction of insecticides with Ha‐Pgp. K_d values of different insecticides for binding to Ha‐Pgp were found to be lower in the presence of cholesterol in the proteoliposomes compared to its absence. Results suggest that cholesterol plays a role in the recognition and interaction of insecticides by modulating Ha‐Pgp ATPase and may be involved in efflux of insecticides from cells by the transporter.