Mukhtasimova_2013_Biophys.J_104_355

Gating of the muscle-type acetylcholine receptor (AChR) channel depends on communication between the ACh-binding site and the remote ion channel. A key region for this communication is located within the structural transition zone between the ligand-binding and pore domains. Here, stemming from beta-strand 10 of the binding domain, the invariant alphaArg209 lodges within the hydrophobic interior of the subunit and is essential for rapid and efficient channel gating. Previous charge-reversal experiments showed that the contribution of alphaArg209 to channel gating depends strongly on alphaGlu45, also within this region. Here we determine whether the contribution of alphaArg209 to channel gating depends on additional anionic or electron-rich residues in this region. Also, to reconcile diverging findings in the literature, we compare the dependence of alphaArg209 on alphaGlu45 in AChRs from different species, and compare the full agonist ACh with the weak agonist choline. Our findings reveal that the contribution of alphaArg209 to channel gating depends on additional nearby electron-rich residues, consistent with both electrostatic and steric contributions. Furthermore, alphaArg209 and alphaGlu45 show a strong interdependence in both human and mouse AChRs, whereas the functional consequences of the mutation alphaE45R depend on the agonist. The emerging picture shows a multifaceted network of interdependent residues that are required for communication between the ligand-binding and pore domains.