Resolving the DDT target protein in insects as a subunit of the ATP synthase.
1,1-bis-(p-Chlorophenyl)-2,2,2-trichloroethane (DDT) inhibited the ATP hydrolytic activity of the ATP synthase from a DDT-susceptible insect (Apis mellifera) as well as a DDT-tolerant insect (Spodoptera littoralis), and from rat liver and bovine heart in a parallel way to its insecticidal properties and selectivity of action. Inhibition of the ATPase activity of these preparations by DDT was parallel to the poisoning of the source organism with DDT. Furthermore, both the inhibition and poisoning of insects were affected similarly by temperature. Inhibition of the insect enzyme activity by DDT was specific and differed from that by oligomycin or N,N-dicyclohexylcarbodi-imide (DCCD). PAGE analysis of the various preparations of the enzyme showed that the inhibition of the enzyme activity by DDT was associated with the presence of a selective protein band with an apparent molecular mass of 23 kDa. This protein band exists in the preparations from the DDT-susceptible insects but was absent from the preparations of the enzyme from the DDT-insensitive sources. Removal of this protein band from the enzyme rendered its activity insensitive to inhibition by DDT. The protein was purified directly from mitochondria and the DDT sensitivity was reconstituted upon its addition to the DDT-insensitive F1-ATPase. We conclude that this identified protein of the ATP synthase is the DDT target protein in insects.
Younis
Hassan M
HM
Laboratory of Bioenergetics and Biochemical Toxicology, Department of Pesticide Chemistry, Faculty of Agriculture, El-Chatby, 21545, Alexandria University, Alexandria, Egypt. hmyounis@dataxprs.com.eg
Abo-El-Saad
Mahmoud M
MM
Abdel-Razik
Reda K
RK
Abo-Seda
Samia A
SA
D000255:Adenosine Triphosphate / Q000378:metabolism; D000818:Animals; D002417:Cattle; D003634:DDT / Q000494:pharmacology*; D004591:Electrophoresis, Polyacrylamide Gel; D004791:Enzyme Inhibitors / Q000494:pharmacology; D006868:Hydrolysis; D007313:Insecta / Q000187:drug effects* / Q000201:enzymology; D007306:Insecticides / Q000494:pharmacology*; D007700:Kinetics; D008297:Male; D008929:Mitochondria, Heart / Q000187:drug effects / Q000201:enzymology; D008930:Mitochondria, Liver / Q000187:drug effects / Q000201:enzymology; D008931:Mitochondria, Muscle / Q000187:drug effects / Q000201:enzymology; D008970:Molecular Weight; D021122:Protein Subunits; D006180:Proton-Translocating ATPases / Q000737:chemistry* / Q000302:isolation & purification / Q000378:metabolism*; D051381:Rats; D013696:Temperature
D004791:Enzyme Inhibitors; D007306:Insecticides; D021122:Protein Subunits; D000255:Adenosine Triphosphate; D003634:DDT; D006180:Proton-Translocating ATPases