title: The Free Energy Landscape of GABA Binding to a Pentameric Ligand-Gated Ion Channel and Its Disruption by Mutations.
abstract: Pentameric ligand-gated ion channels (pLGICs) of the Cys-loop superfamily are important neuroreceptors that mediate fast synaptic transmission. They are activated by the binding of a neurotransmitter, but the details of this process are still not fully understood. As a prototypical pLGIC, here we choose the insect resistance to dieldrin (RDL) receptor involved in resistance to insecticides and investigate the binding of the neurotransmitter GABA to its extracellular domain at the atomistic level. We achieve this by means of μ-sec funnel-metadynamics simulations, which efficiently enhance the sampling of bound and unbound states by using a funnel-shaped restraining potential to limit the exploration in the solvent. We reveal the sequence of events in the binding process from the capture of GABA from the solvent to its pinning between the charged residues Arg111 and Glu204 in the binding pocket. We characterize the associated free energy landscapes in the wild-type RDL receptor and in two mutant forms, where the key residues Arg111 and Glu204 are mutated to Ala. Experimentally these mutations produce nonfunctional channels, which is reflected in the reduced ligand binding affinities due to the loss of essential interactions. We also analyze the dynamical behavior of the crucial loop C, whose opening allows the access of GABA to the binding site and closure locks the ligand into the protein. The RDL receptor shares structural and functional features with other pLGICs; hence, our work outlines a valuable protocol to study the binding of ligands to pLGICs beyond conventional docking and molecular dynamics techniques.
authors.affiliation: Department of Physics, King's College London , Strand, London WC2R 2LS, United Kingdom.
authors.forename: Federico
authors.initials: F
authors.lastname: Comitani
authors.affiliation: Universitá della Svizzera Italiana (USI) , Faculty of Informatics, Institute of Computational Science - Center for Computational Medicine in Cardiology, via G. Buffi 13, CH-6900 Lugano, Switzerland.|Department of Pharmacy, University of Naples "Federico II" , via D. Montesano 49, I-80131 Naples, Italy.
authors.forename: Vittorio
authors.initials: V
authors.lastname: Limongelli
authors.affiliation: Department of Physics, King's College London , Strand, London WC2R 2LS, United Kingdom.
authors.forename: Carla
authors.initials: C
authors.lastname: Molteni
mesh_terms: D001665:Binding Sites; D058446:Ligand-Gated Ion Channels / Q000737:chemistry*; D056004:Molecular Dynamics Simulation; D009154:Mutation; D005680:gamma-Aminobutyric Acid / Q000737:chemistry*
chemical_list: D058446:Ligand-Gated Ion Channels; D005680:gamma-Aminobutyric Acid