α-Conotoxin AuIB Selectively Blocks α3β4 Nicotinic Acetylcholine Receptors and Nicotine-Evoked Norepinephrine Release

Neuronal nicotinic acetylcholine receptors (nAChRs) with putative α3β4-subunits have been implicated in the mediation of signaling in various systems, including ganglionic transmission peripherally and nicotine-evoked neurotransmitter release centrally. However, progress in the characterization of these receptors has been hampered by a lack of α3β4-selective ligands. In this report, we describe the purification and characterization of an α3β4 nAChR antagonist, α-conotoxin AuIB, from the venom of the “court cone,”Conus aulicus. We also describe the total chemical synthesis of this and two related peptides that were also isolated from the venom. α-Conotoxin AuIB blocks α3β4 nAChRs expressed in Xenopus oocytes with an IC50 of 0.75 μm, a kon of 1.4 × 106 min-1m−1, a koffof 0.48 min-1, and aKd of 0.5 μm. Furthermore, α-conotoxin AuIB blocks the α3β4 receptor with >100-fold higher potency than other receptor subunit combinations, including α2β2, α2β4, α3β2, α4β2, α4β4, and α1β1γδ. Thus, AuIB is a novel, selective probe for α3β4 nAChRs. AuIB (1–5 μm) blocks 20–35% of the nicotine-stimulated norepinephrine release from rat hippocampal synaptosomes, whereas nicotine-evoked dopamine release from striatal synaptosomes is not affected. Conversely, the α3β2-specific α-conotoxin MII (100 nm) blocks 33% of striatal dopamine release but not hippocampal norepinephrine release. This suggests that in the respective systems, α3β4-containing nAChRs mediate norepinephrine release, whereas α3β2-containing receptors mediate dopamine release.