Subunit symmetry at the extracellular domain-transmembrane domain interface in acetylcholine receptor channel gating.

Transmitter molecules bind to synaptic acetylcholine receptor channels (AChRs) to promote a global channel-opening conformational change. Although the detailed mechanism that links ligand binding and channel gating is uncertain, the energy changes caused by mutations appear to be more symmetrical between subunits in the transmembrane domain compared with the extracellular domain. The only covalent connection between these domains is the pre-M1 linker, a stretch of five amino acids that joins strand β10 with the M1 helix. In each subunit, this linker has a central Arg (Arg(3')), which only in the non-α-subunits is flanked by positively charged residues. Previous studies showed that mutations of Arg(3') in the α-subunit alter the gating equilibrium constant and reduce channel expression. We recorded single-channel currents and estimated the gating rate and equilibrium constants of adult mouse AChRs with mutations at the pre-M1 linker and the nearby residue Glu(45) in non-α-subunits. In all subunits, mutations of Arg(3') had similar effects as in the α-subunit. In the ε-subunit, mutations of the flanking residues and Glu(45) had only small effects, and there was no energy coupling between εGlu(45) and εArg(3'). The non-α-subunit Arg(3') residues had Φ-values that were similar to those for the α-subunit. The results suggest that there is a general symmetry between the AChR subunits during gating isomerization in this linker and that the central Arg is involved in expression more so than gating. The energy transfer through the AChR during gating appears to mainly involve Glu(45), but only in the α-subunits.