Tricyclic antidepressants and mecamylamine bind to different sites in the human α4β2 nicotinic receptor ion channel

The interaction of tricyclic antidepressants with the human (h) α4β2 nicotinic acetylcholine receptor in different conformational states was compared with that for the noncompetitive antagonist mecamylamine by using functional and structural approaches. The results established that: (a) [3H]imipramine binds to hα4β2 receptors with relatively high affinity (Kd = 0.83 ± 0.08 μM), but imipramine does not differentiate between the desensitized and resting states, (b) although tricyclic antidepressants inhibit (±)-epibatidine-induced Ca2+ influx in HEK293-hα4β2 cells with potencies that are in the same concentration range as that for (±)-mecamylamine, tricyclic antidepressants inhibit [3H]imipramine binding to hα4β2 receptors with affinities >100-fold higher than that for (±)-mecamylamine. This can be explained by our docking results where imipramine interacts with the leucine (position 9′) and valine (position 13′) rings by van der Waals contacts, whereas mecamylamine interacts electrostatically with the outer ring (position 20′), (c) van der Waals interactions are in agreement with the thermodynamic results, indicating that imipramine interacts with the desensitized and resting receptors by a combination of enthalpic and entropic components. However, the entropic component is more important in the desensitized state, suggesting local conformational changes. In conclusion, our data indicate that tricyclic antidepressants and mecamylamine efficiently inhibit the ion channel by interacting at different luminal sites. The high proportion of protonated mecamylamine calculated at physiological pH suggests that this drug can be attracted to the channel mouth before binding deeper within the receptor ion channel finally blocking ion flux.