Identification of Critical Elements Determining Toxins and Insecticide Affinity, Ligand Binding Domains and Channel Properties

  • Hélène Tricoire-Leignel
  • Steeve Hervé ThanyEmail author
Part of the Advances in Experimental Medicine and Biology book series (AEMB, volume 683)


Insect nicotinic acetylcholine receptors have been objects of attention since the discovery of neonicotinoid insecticides. Mutagenesis studies have revealed that, although the detailed subunit composition of insect nicotinic acetylcholine receptors subtypes eludes us, the framework provided by mutagenesis analysis makes a picture of the subunits involved in the ligand binding and channel properties. In fact, many residues that line the channel or bind to the ligand seemed to be strongly conserved in particular in the N-terminal extracellular region and the second transmembrane domain which constitutes the ion-conducting pathway supporting the flux of ions as well as their discrimination. In fact, the positions are carried by loops B and C, respectively, which contain amino acids directly contributing to the acetylcholine binding site. Mutation of these residues accounts for insect resistance to neonicotinoid insecticides such as imidacloprid or a loss of specific binding. The discovery of the same mutation at homologous residues in different insect species or its conservation raises the intriguing question of whether a single mutation is essential to generate a resistance phenotype or whether some subunit confer insensitivity to ligand. Consequently, recent finding using information from Torpedo marmorata α1 subunit and soluble Aplysia californica and Lymnae stagnalis acetylcholine binding proteins from crystallization suggest that insect nAChR subunits had contributing amino acids in the agonist site structure which participate to affinity and pharmacological properties of these receptors. These new range of data greatly facilitate the understanding of toxin-nAChR interactions and the neonicotinoid binding and selectivity.


Nicotinic Receptor Nicotinic Acetylcholine Receptor nAChR Subtype Neuronal Nicotinic Acetylcholine Receptor Neuronal Nicotinic Receptor 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.


Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.


  1. 1.
    Jones AK, Sattelle DB. The cys-loop ligand-gated ion channel superfamily of the honeybee, Apis mellifera. Invert Neurosci 2006; 6(3):123–132.PubMedCrossRefGoogle Scholar
  2. 2.
    Jones AK, Sattelle DB. The cys-loop ligand-gated ion channel gene superfamily of the red flour beetle, Tribolium castaneum. BMC Genomics 2007; 8:327.PubMedCrossRefGoogle Scholar
  3. 3.
    Lansdell SJ, Millar NS. Cloning and heterologous expression of Dalpha4, a drosophila neuronal nicotinic acetylcholine receptor subunit: Identification of an alternative exon influencing the efficiency of subunit assembly. Neuropharmacology 2000; 39(13):2604–2614.PubMedCrossRefGoogle Scholar
  4. 4.
    Grauso M, Reenan RA, Culetto E et al. Novel putative nicotinic acetylcholine receptor subunit genes, Dalpha5, Dalpha6 and Dalpha7, in Drosophila melanogaster identify a new and highly conserved target of adenosine deaminase acting on RNA-mediated A-to-I premrna editing. Genetics 2002; 160(4):1519–1533.PubMedGoogle Scholar
  5. 5.
    Sattelle DB, Jones AK, Sattelle BM et al. Edit, cut and paste in the nicotinic acetylcholine receptor gene family of Drosophila melanogaster. Bioessays 2005; 27(4):366–376.PubMedCrossRefGoogle Scholar
  6. 6.
    Jones AK, Buckingham SD, Brown LA et al. Alternative splicing of the Anopheles gambiae nicotinic acetylcholine receptor, Agam alphabeta9, generates both α and β subunits. Invert Neurosci 2009; 9(2):77–84.PubMedCrossRefGoogle Scholar
  7. 7.
    Arias HR. Localization of agonist and competitive antagonist binding sites on nicotinic acetylcholine receptors. Neurochem Int 2000; 36(7):595–645.PubMedCrossRefGoogle Scholar
  8. 8.
    Corringer PJ, Le Novere N, Changeux JP. Nicotinic receptors at the amino acid level. Annu Rev Pharmacol Toxicol 2000; 40:431–458.PubMedCrossRefGoogle Scholar
  9. 9.
    Unwin N. Projection structure of the nicotinic acetylcholine receptor: Distinct conformations of the α subunits. J Mol Biol 1996; 257(3):586–596.PubMedCrossRefGoogle Scholar
  10. 10.
    Brejc K, van Dijk WJ, Klaassen RV et al. Crystal structure of an ACh-Binding Protein reveals the ligand-binding domain of nicotinic receptors. Nature 2001; 411(6835):269–276.PubMedCrossRefGoogle Scholar
  11. 11.
    Smit AB, Syed NI, Schaap D et al. A glia-derived acetylcholine-binding protein that modulates synaptic transmission. Nature 2001; 411(6835):261–268.PubMedCrossRefGoogle Scholar
  12. 12.
    Hansen SB, Talley TT, Radic Z et al. Structural and ligand recognition characteristics of an AcetylCholine-Binding Protein from Aplysia californica. J Biol Chem 2004; 279(23):24197–24202.PubMedCrossRefGoogle Scholar
  13. 13.
    Celie PH, van Rossum-Fikkert SE, van Dijk WJ et al. Nicotine and carbamylcholine binding to nicotinic acetylcholine receptors as studied in AChBP crystal structures. Neuron 2004; 41(6):907–914.PubMedCrossRefGoogle Scholar
  14. 14.
    Hibbs RE, Talley TT, Taylor P. Acrylodan-conjugated cysteine side chains reveal conformational state and ligand site locations of the acetylcholine-binding protein. J Biol Chem 2004; 279(27):28483–28491.PubMedCrossRefGoogle Scholar
  15. 15.
    Sine SM. The nicotinic receptor ligand binding domain. J Neurobiol 2002; 53(4):431–446.PubMedCrossRefGoogle Scholar
  16. 16.
    Tomizawa M, Casida JE. Molecular recognition of neonicotinoid insecticides: The determinants of life or death. Acc Chem Res 2009; 42(2):260–269.PubMedCrossRefGoogle Scholar
  17. 17.
    Tsetlin VI, Hucho F. Snake and snail toxins acting on nicotinic acetylcholine receptors: Fundamental aspects and medical applications. FEBS Lett 2004; 557(1–3):9–13.PubMedCrossRefGoogle Scholar
  18. 18.
    Dutertre S, Lewis RJ. Computational approaches to understand alpha-conotoxin interactions at neuronal nicotinic receptors. Eur J Biochem 2004; 271(12):2327–2334.PubMedCrossRefGoogle Scholar
  19. 19.
    Ulens C, Hogg RC, Celie PH et al. Structural determinants of selective α-conotoxin binding to a nicotinic acetylcholine receptor homolog AChBP. Proc Natl Acad Sci USA 2006; 103(10):3615–3620.PubMedCrossRefGoogle Scholar
  20. 20.
    Dutertre S, Ulens C, Buttner R et al. AChBP-targeted α-conotoxin correlates distinct binding orientations with nAChR subtype selectivity. EMBO J 2007; 26(16):3858–3867.PubMedCrossRefGoogle Scholar
  21. 21.
    Bourne Y, Talley TT, Hansen SB et al. Crystal structure of a Cbtx-AChBP complex reveals essential interactions between snake alpha-neurotoxins and nicotinic receptors. EMBO J 2005; 24(8):1512–1522.PubMedCrossRefGoogle Scholar
  22. 22.
    Terlau H, Olivera BM. Conus venoms: A rich source of novel ion channel-targeted peptides. Physiol Rev 2004; 84(1):41–68.CrossRefGoogle Scholar
  23. 23.
    Janes RW. Alpha-conotoxins as selective probes for nicotinic acetylcholine receptor subclasses. Curr Opin Pharmacol 2005; 5(3):280–292.PubMedCrossRefGoogle Scholar
  24. 24.
    Matsuda K, Shimomura M, Kondo Y et al. Role of loop D of the α7 nicotinic acetylcholine receptor in its interaction with the insecticide imidacloprid and related neonicotinoids. Br J Pharmacol 2000; 130(5):981–986.PubMedCrossRefGoogle Scholar
  25. 25.
    Shimomura M, Satoh H, Yokota M et al. Insect-vertebrate chimeric nicotinic acetylcholine receptors identify a region, loop B to the N-terminus of the drosophila Dα2 subunit, which contributes to neonicotinoid sensitivity. Neurosci Lett 2005; 385(2):168–172.PubMedCrossRefGoogle Scholar
  26. 26.
    Shimomura M, Yokota M, Matsuda K et al. Roles of loop C and the loop B-C interval of the nicotinic receptor alpha subunit in its selective interactions with imidacloprid in insects. Neurosci Lett 2004; 363(3):195–198.PubMedCrossRefGoogle Scholar
  27. 27.
    Shimomura M, Yokota M, Ihara M et al. Role in the selectivity of neonicotinoids of insect-specific basic residues in loop D of the nicotinic acetylcholine receptor agonist binding site. Mol Pharmacol 2006; 70(4):1255–1263.PubMedCrossRefGoogle Scholar
  28. 28.
    Yao X, Song F, Chen F et al. Amino acids within loops D, E and F of insect nicotinic acetylcholine receptor beta subunits influence neonicotinoid selectivity. Insect Biochem Mol Biol 2008; 38(9):834–840.PubMedCrossRefGoogle Scholar
  29. 29.
    Toshima K, Kanaoka S, Yamada A et al. Combined roles of loops C and D in the interactions of a neonicotinoid insecticide imidacloprid with the α4β2 nicotinic acetylcholine receptor. Neuropharmacology 2009; 56(1):264–272.PubMedCrossRefGoogle Scholar
  30. 30.
    Jeschke P, Nauen R. Neonicotinoids-from zero to hero in insecticide chemistry. Pest Manag Sci 2008; 64(11):1084–1098.PubMedCrossRefGoogle Scholar
  31. 31.
    Liu Z, Williamson MS, Lansdell SJ et al. A nicotinic acetylcholine receptor mutation conferring target-site resistance to imidacloprid in Nilaparvata lugens (brown planthopper). Proc Natl Acad Sci USA 2005; 102(24):8420–8425.PubMedCrossRefGoogle Scholar
  32. 32.
    Wang Y, Cheng J, Qian X et al. Actions between neonicotinoids and key residues of insect nAChR based on an ab initio quantum chemistry study: Hydrogen bonding and cooperative pi-pi interaction. Bioorg Med Chem 2007; 15(7):2624–2630.PubMedCrossRefGoogle Scholar
  33. 33.
    Ihara M, Okajima T, Yamashita A et al. Crystal structures of Lymnaea stagnalis AChBP in complex with neonicotinoid insecticides imidacloprid and clothianidin. Invert Neurosci 2008; 8(2):71–81.PubMedCrossRefGoogle Scholar
  34. 34.
    Talley TT, Harel M, Hibbs RE et al. Atomic interactions of neonicotinoid agonists with AChBP: Molecular recognition of the distinctive electronegative pharmacophore. Proc Natl Acad Sci USA 2008; 105(21):7606–7611.PubMedCrossRefGoogle Scholar
  35. 35.
    Ohno I, Tomizawa M, Durkin KA et al. Molecular features of neonicotinoid pharmacophore variants interacting with the insect nicotinic receptor. Chem Res Toxicol 2009.Google Scholar
  36. 36.
    Ohno I, Tomizawa M, Durkin KA et al. Bis-neonicotinoid insecticides: Observed and predicted binding interactions with the nicotinic receptor. Bioorg Med Chem Lett 2009; 19(13):3449–3452.PubMedCrossRefGoogle Scholar
  37. 37.
    Rocher A, Marchand-Geneste N. Homology modelling of the Apis mellifera nicotinic acetylcholine receptor (nAChR) and docking of imidacloprid and fipronil insecticides and their metabolites. SAR QSAR Environ Res 2008; 19(3–4):245–261.PubMedCrossRefGoogle Scholar
  38. 38.
    Galzi JL, Devillers-Thiery A, Hussy N et al. Mutations in the channel domain of a neuronal nicotinic receptor convert ion selectivity from cationic to anionic. Nature 1992; 359(6395):500–505.PubMedCrossRefGoogle Scholar
  39. 39.
    Corringer PJ, Bertrand S, Bohler S et al. Critical elements determining diversity in agonist binding and desensitization of neuronal nicotinic acetylcholine receptors. J Neurosci 1998; 18(2):648–657.PubMedGoogle Scholar
  40. 40.
    Imoto K, Busch C, Sakmann B et al. Rings of negatively charged amino acids determine the acetylcholine receptor channel conductance. Nature 1988; 335(6191):645–648.PubMedCrossRefGoogle Scholar
  41. 41.
    Jensen AA, Frolund B, Liljefors T et al. Neuronal nicotinic acetylcholine receptors: Structural revelations, target identifications and therapeutic inspirations. J Med Chem 2005; 48(15):4705–4745.PubMedCrossRefGoogle Scholar
  42. 42.
    Buckingham SD, Adcock C, Sansom MS et al. Functional characterization of a mutated chicken alpha7 nicotinic acetylcholine receptor subunit with a leucine residue inserted in transmembrane domain 2. Br J Pharmacol 1998; 124(4):747–755.PubMedCrossRefGoogle Scholar
  43. 43.
    Bertrand D, Devillers-Thiery A, Revah F et al. Unconventional pharmacology of a neuronal nicotinic receptor mutated in the channel domain. Proc Natl Acad Sci USA 1992; 89(4):1261–1265.PubMedCrossRefGoogle Scholar
  44. 44.
    Palma E, Mileo AM, Eusebi F et al. Threonine-for-leucine mutation within domain M2 of the neuronal alpha(7) nicotinic receptor converts 5-hydroxytryptamine from antagonist to agonist. Proc Natl Acad Sci USA 1996; 93(20):11231–11235.PubMedCrossRefGoogle Scholar
  45. 45.
    Palma E, Fucile S, Barabino B et al. Strychnine activates neuronal alpha7 nicotinic receptors after mutations in the leucine ring and transmitter binding site domains. Proc Natl Acad Sci USA 1999; 96(23):13421–13426.PubMedCrossRefGoogle Scholar
  46. 46.
    Courjaret R, Lapied B. Complex intracellular messenger pathways regulate one type of neuronal alpha-bungarotoxin-resistant nicotinic acetylcholine receptors expressed in insect neurosecretory cells (Dorsal Unpaired Median neurons). Mol Pharmacol 2001; 60(1):80–91.PubMedGoogle Scholar

Copyright information

© Landes Bioscience and Springer Science+Business Media 2010

Authors and Affiliations

  • Hélène Tricoire-Leignel
    • 1
  • Steeve Hervé Thany
    • 1
    Email author
  1. 1.Laboratoire Récepteurs et Canaux Ioniques Membranaires (RCIM), UPRES EA 2647/USC INRA 2023, IFR 149 QUASAVUniversité d’Angers, UFR de SciencesAngersFrance

Personalised recommendations