Molecular simulation study of the unbinding of α-conotoxin [ϒ4E]GID at the α7 and α4β2 neuronal nicotinic acetylcholine receptors.

The α7 and α4β2 neuronal nicotinic receptors belonging to the family of ligand-gated ion channels are most prevalent in the brain, and are implicated in various neurodegenerative disorders. α-conotoxin GID (and its analogue [ϒ4E]GID) specifically inhibits these subtypes, with more affinity towards the human α7 (hα7) subtype, and is valuable in understanding the physiological roles of these receptors. In this study, we use umbrella-sampling molecular dynamics simulations to understand the mechanism of interaction between [ϒ4E]GID and the agonist binding pockets of the α4β2 and the hα7 receptors, and to estimate their relative binding affinities (ΔG). The obtained ΔG values indicate stronger interaction with the hα7 receptor, in agreement with previous experimental studies. Simulations also revealed different unbinding pathways between the two receptor subtypes, enabling identification of a number of interactions at locations far from the orthosteric binding site which may explain the difference in [ϒ4E]GID potency. The pathways identified will help in the design of novel conotoxins with increased potency at α4β2, for which there is currently no known highly potent conotoxin inhibitor. Computational mutational free energy analyses also revealed a number of possible single-site mutations to GID which might enhance its selective binding to α4β2 over α7.