β2 Subunit Contribution to 4/7 α-Conotoxin Binding to the Nicotinic Acetylcholine Receptor - Archive ouverte HAL Accéder directement au contenu
Article Dans Une Revue Journal of Biological Chemistry Année : 2005

β2 Subunit Contribution to 4/7 α-Conotoxin Binding to the Nicotinic Acetylcholine Receptor

Résumé

The structures of acetylcholine-binding protein (AChBP) and nicotinic acetylcholine receptor (nAChR) homology models have been used to interpret data from mutagenesis experiments at the nAChR. However, little is known about AChBP-derived structures as predictive tools. Molecular surface analysis of nAChR models has revealed a conserved cleft as the likely binding site for the 4/7 α-conotoxins. Here, we used an α3β2 model to identify β2 subunit residues in this cleft and investigated their influence on the binding of α-conotoxins MII, PnIA, and GID to the α3β2 nAChR by two-electrode voltage clamp analysis. Although a β2-L119Q mutation strongly reduced the affinity of all three α-conotoxins, β2-F117A, β2-V109A, and β2-V109G mutations selectively enhanced the binding of MII and GID. An increased activity of α-conotoxins GID and MII was also observed when the β2-F117A mutant was combined with the α4 instead of the α3 subunit. Investigation of A10L-PnIA indicated that high affinity binding to β2-F117A, β2-V109A, and β2-V109G mutants was conferred by amino acids with a long side chain in position 10 (PnIA numbering). Docking simulations of 4/7 α-conotoxin binding to the α3β2 model supported a direct interaction between mutated nAChR residues and α-conotoxin residues 6, 7, and 10. Taken together, these data provide evidence that the β subunit contributes to α-conotoxin binding and selectivity and demonstrate that a small cleft leading to the agonist binding site is targeted by α-conotoxins to block the nAChR.
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Dates et versions

hal-02306844 , version 1 (07-10-2019)

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Sébastien Dutertre, Annette Nicke, Richard J Lewis. β2 Subunit Contribution to 4/7 α-Conotoxin Binding to the Nicotinic Acetylcholine Receptor. Journal of Biological Chemistry, 2005, 280 (34), pp.30460-30468. ⟨10.1074/jbc.M504229200⟩. ⟨hal-02306844⟩
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