Acetylcholinesterase and Insecticide Resistance

Physical models of acetylcholinesterase are used to explore a wide range of topics, including enzyme, active site, catalytic site, substrate and inhibitor.  Similar to the serine proteases, acetylcholinesterase has a catalytic triad composed of serine, histidine, and a negatively charged amino acid.  (Acetylcholinesterase uses glutamine, whereas serine proteases use aspartate.)  The catalytic site of acetylcholinesterase is situated deep within a narrow active site gorge, yet the reaction rate is one of the fastest known, close to the rate of diffusion.  Acetylcholinesterase is the target of organophosphate and carbamide insecticides; a point mutation in the enzyme renders it resistant to these insecticides.  Students can explore this molecular story using physical models, jmol tutorials and paper bioinformatics.

:: Acetylcholinesterase active site model with substrate, inhibitor and mutation

:: Jmol tutorials

                       
:: Bioinformatics activity exploring insecticide resistance ‘Evolution in Action’

:: Molecule of the Month extension

 

Acetylcholinesterase References
Bigbee, John W., Sharma, Karun V., Gupta, Jyotsna J., and Dupree, Jeffrey L. 1999. Morphogenic Role for Acetylcholinesterase in Axonal Outgrowth During Neural Development. Environmental Health Perspectives Supplements 107(S1):81-87.

De Ferrari, G.V., Canales, M.A., Shin, I., Weiner, L.M., Silman, I. and Inestrosa, N.C. 2001. A structural motif of acetylcholinesterase that promotes amyloid beta-peptide fibril formation. Biochem. 40(35):10447-10457.

Inestrosa, Nibaldo C., Alejandra Alvarez, Cristián A. Pérez, Ricardo D. Moreno, Matias Vicente, Claudia Linker, Olivia I. Casanueva, Claudio Soto, and Jorge Garrido. 1996. Acetylcholinesterase Accelerates Assembly of Amyloid-β-Peptides into Alzheimer’s Fibrils: Possible Role of the Peripheral Site of the Enzyme. Cell 16:881-891. 

Szegletes, Tivadar, Mallender, William D., Thomas, Patrick J. and Rosenberry, Terrone L. 1999. Substrate binding to the peripheral site of acetylcholinesterase initiates enzymatic catalysis. Substrate inhibition arises as a secondary effect. Biochem. 38:122-133.

 

References pertaining to alternate splice sites and their function
Bartels, Cynthia F., Zelinski, Teresa, and Lockridge, Oksana. 1993. Mutation at codon 322 in the human acetylcholinesterase (ACHE) gene accounts for YT blood group polymorphism. Am. J. Genet. 52:928-936.

Dvir, Hay., Harel, Michal, Bon, Suzanne, Liu, Wang-Qing, Vidal, Michel, Garbay, Christiane, Sussman, Joel L., Massoulié and Silman, Israel. 2004. The synaptic acetylcholinesterase tetramer assembles around a polyproline II helix. EMBO J. 23:4394-4405.

Li, Ying, Camp, Shelley, Rachinsky, Tara L., Getman, Damon and Taylor, Palmer. 1991. Gene Structure of Mammalian Acetylcholinesterase: Alternative Exons Dictate Tissue-Specific Expression. J. Biol. Chem. 34: 23083-23090.

Massoulié, Jean, Bon, Suzanne, Perrier, Noël, and Falasca Cinzia. 2005. The C-terminal peptides of acetylcholinesterase: Cellular trafficking, oligomerization and functional anchoring. Chemico-Biological Interactions 157-158:3-14.

Massoulié, Jean, Anselmet, Alain, Bon, Suzanne, Krejci, Eric, Legay, Clair, Morel, Nathalie and Simon, Stéphanie. 1999. The polymorphism of acetylcholinesterase: post-translational processing, quaternary associations and localization. Chemico-Biological Interactions 119-120:29-42.

Perrier, Anselme L., Massoulié, Jean, and Krejci, Eric. 2002. PRiMA: The Membrane Anchor of Acetylcholinesterase in the Brain. Neuron 33:275-285.

Schumacher, Mark, Maulet, Yves, Camp, Shelley and Taylor, Palmer. 1988. Multiple Messenger RNA Species Give Rise to the Structural Diversity in Acetylcholinesterase. J. Biol. Chem. 263(35):18879-18987.

Sternfeld, Meira, Shoham, Shai, Klein, Omer, Flores-Flores, Cesar, Evron, Tamah, Idelson, Gregory H., Kitsberg, Dani, Patrick, James W. and Soreq, Hermona. 2000. Excess "read-through" acetylcholinesterase attenuates but the "synaptic" variant intensifies neurodeterioration correlates. Proc. Natl. Acad. Sci. 97(15):8647-8652.

 

References pertaining insecticide resistance: fitness cost and population genetics
Berticat, Claire, Boquien, Grégoire, Raymond, Michel and Chevillon, Christine. 2002. Insecticide resistance genes induce a mating competition cost in Culex pipiens mosquitoes. Genet. Res. Camb. 79:41-47.
Berticat, C., Duron, O., Heyse, D. and Raymond, M. 2004. Insecticide resistance genes confer a predation cost on mosquitoes, Culex pipiens. Genet. Res., Camb. 83:189-196.
Berticat, Claire, Rousset, François, Raymond, Michel, Berthomieu, Arnaud and Weill, Mylène. 2002. High Wolbachia density in insecticide-resistant mosquitoes. Proc. Royal Soc. London B 269(1498):1413-1416.
Bourguet, Denis, Guillemaud, Thomas, Chevillon, Christine, and Raymond, Michel. 2004. Fitness costs of insecticide resistance in natural breeding sites of the mosquito Culex pipiens. Evol. 58(!):128-135.
Chevillon, Christine, Raymond, Michel, Guillemaud, Thomas, Lenormand, Thomas and Pasteur, Nicole. 1999. Population genetics of insecticide resistance in the mosquito Culex pipiens. Biol. J. Linnean Soc. 69:147-157.
Eritja, R and Chevillon, C.1999. Interruption of chemical mosquito control and evolution of insecticide resistance genes in Culex pipiens (Diptera: Culicidae). J. Med. Entomol. 36(1):41-49.
Gazave, Élodie, Chevillon, Christine, Lenormand, Thomas, Marquine, Maïté and Raymond, Michel. 2001. Dissecting the cost of insecticide resistance genes during the overwintering period of the mosquito. Heredity 87(4):441-448.
Labbe, P. Lenormand, T. and Raymond, M. 2005. On the worldwide spread of an insecticide resistance gene: a role for local selection. J. Evol. Biol. 18:1471-1484.
Lenormand, Thomas, Bourguet, Denis, Guillemaud, Thomas, and Raymond, Michel. 1999. Tracking the evolution of insecticide resistance in the mosquito Culex pipiens. Nature 400(26 Aug):861-864.
Lenormand, Thomas, Guillemaud, Thomas, Bourguet, Denis and Raymond, Michel. 1998. Evaluating Gene Flow Using Selected Markers: A Case Study. Genetics 149:1383-1392.
Raymond, M., Berticat, C., Weill, M., Pasteur, N. and Chevillon, C. 2001. Insecticide resistance in the mosquito Culex pipiens: what have we learned about adaptation. Genetica 112-113:287-296.
Shi, Ming An, Lougarre, Andrée, Alies, Carole, Isabelle Frémau, Tang, Zhen Hue, Stojan, Jure, and Fournier, Didier. 2004. Acetylcholinesterase alterations reveal the fitness cost of mutations conferring insecticide resistance. BMC Evolutionary Biology 2004, 4:5