Existence of two acetylcholinesterases in the mosquito Culex pipiens (Diptera:Culicidae).
Two acetylcholinesterases (AChEs), AChE1 and AChE2, differing in substrate specificity and in some aspects of inhibitor sensitivity, have been characterized in the mosquito Culex pipiens. The results of ultracentrifugation in sucrose gradients and nondenaturing gel electrophoresis of AChE activity peak fractions show that each AChE is present as two molecular forms: one amphiphilic dimer possessing a glycolipid anchor and one hydrophilic dimer that does not interact with nondenaturing detergents. Treatment by phosphatidylinositol-specific phospholipase C converts each type of amphiphilic dimer into the corresponding hydrophilic dimer. Molecular forms of AChE1 have a lower electrophoretic mobility than those of AChE2. However, amphiphilic dimers and hydrophilic dimers have similar sedimentation coefficients (5.5S and 6.5S, respectively). AChE1 and AChE2 dimers, amphiphilic or hydrophilic, resist dithiothreitol reduction under conditions that allow reduction of Drosophila AChE dimers. In the insecticide-susceptible strain S-LAB, AChE1 is inhibited by 5 x 10(-4) M propoxur (a carbamate insecticide), whereas AChE2 is resistant. All animals are killed by this concentration of propoxur, indicating that only AChE1 fulfills the physiological function of neurotransmitter hydrolysis at synapses. In the insecticide-resistant strain, MSE, there is no mortality after exposure to 5 x 10(-4) M propoxur: AChE2 sensitivity to propoxur is unchanged, whereas AChE1 is now resistant to 5 x 10(-4) M propoxur. The possibility that AChE1 and AChE2 are products of tissue-specific posttranslational modifications of a single gene is discussed, but we suggest, based on recent results obtained at the molecular level in mosquitoes, that they are encoded by two different genes.
Bourguet
D
D
Laboratoire Génétique et Environnement, Université Montpellier-II, France.
Raymond
M
M
Fournier
D
D
Malcolm
C A
CA
Toutant
J P
JP
Arpagaus
M
M
D000110:Acetylcholinesterase / Q000302:isolation & purification / Q000378:metabolism*; D000818:Animals; D002800:Cholinesterase Inhibitors / Q000494:pharmacology*; D003465:Culex / Q000201:enzymology*; D004198:Disease Susceptibility; D004229:Dithiothreitol / Q000494:pharmacology; D004591:Electrophoresis, Polyacrylamide Gel; D017261:Glycosylphosphatidylinositols / Q000032:analysis; D007527:Isoenzymes / Q000302:isolation & purification / Q000378:metabolism*; D007814:Larva; D008970:Molecular Weight; D009439:Neuraminidase; D001074:Propoxur / Q000494:pharmacology; D013045:Species Specificity; D013379:Substrate Specificity; D014461:Ultracentrifugation
D002800:Cholinesterase Inhibitors; D017261:Glycosylphosphatidylinositols; D007527:Isoenzymes; D001074:Propoxur; D000110:Acetylcholinesterase; D009439:Neuraminidase; D004229:Dithiothreitol