Pentameric ligand-gated ion channels (pLGICs) mediate fast chemoelectrical transduction in the nervous system. The mechanism by which the energy of ligand binding leads to current-conducting receptors is poorly understood and may vary among family members. We addressed these questions by correlating the structural and energetic mechanisms by which a naturally occurring M1 domain mutation (α1(Q-26'E)) enhances receptor activation in homo- and heteromeric glycine receptors. We systematically altered the charge of spatially clustered residues at positions 19' and 24', in the M2 and M2-M3 linker domains, respectively, which are known to be critical to efficient receptor activation, on a background of α1(Q-26'E). Changes in the durations of single receptor activations (clusters) and conductance were used to determine interaction coupling energies, which we correlated with conformational displacements as measured in pLGIC crystal structures. Presence of the α1(Q-26'E) enhanced cluster durations and reduced channel conductance in homo- and heteromeric receptors. Strong coupling between α1(-26') and α1(19') across the subunit interface suggests an important role in receptor activation. A lack of coupling between α1(-26') and α1(24') implies that 24' mutations disrupt activation via other interactions. A similar lack of energetic coupling between α1(-26') and reciprocal mutations in the β subunit suggests that this subunit remains relatively static during receptor activation. However, the channel effects of α1(Q-26'E) on α1β receptors suggests at least one α1-α1 interface per pentamer. The coupling-energy change between α1(-26') and α1(19') correlates with a local structural rearrangement essential for pLGIC activation, implying it comprises a key energetic pathway in activating glycine receptors and other pLGICs.
Keywords: Chloride Channel; Cys-loop Receptor; Energetics; Ion Channel; Pentameric Ligand-gated Ion Channel; Receptor Structure-function; Single Channel Kinetics.
© 2015 by The American Society for Biochemistry and Molecular Biology, Inc.