Molecular Recognition Properties of Nicotinic Ligands Determining Selectivity Between Insect and Mammalian Receptors

This investigation defines the roles of various amino acids, neighboring key conserved amino acids in loops C and D of the nicotinic acetylcholine (ACh) receptor (nAChR), in the selective molecular recognition of nicotinic ligands with diverse pharmacophores using Aplysia californica ACh binding protein Y55W (Ac-AChBP) mutants (+Q57R; + Q57R+S189 V; + Q57R+S189E; + Q57T; + Q57T+S189 V; + Q57T+S189E) and Lymnaea stagnalis AChBP (Ls-AChBP) mutants (Q55T; Q55T+S186E; Q55R) as insect and mammalian nAChR structural surrogates, respectively. N-nitro/cyanoimine insecticides show high affinity to four Ac-AChBPs containing Arg57 or Thr57 and Ser189 or Val189, except for those with Glu189. Pyrazinoyl compound selectively interacts with the three Ac-AChBPs containing Arg57 and Ser189, Val189, or Glu189. Cationic ligands prefer three Ac-AChBPs with Thr57 and Ser189, Val189, or Glu189 and two Ls-AChBPs providing Thr55 ± Glu186 over the four Ac- and Ls-AChBPs with Arg57/55. Accordingly, loop C contributes to N-nitro/cyanoimine insecticide action, and loop D controls the affinity of the pyrazinoyl or cationic ligand.