Characterizing the binding and function of TARP γ8-selective AMPA receptor modulators

J Biol Chem. 2020 Oct 23;295(43):14565-14577. doi: 10.1074/jbc.RA120.014135. Epub 2020 Aug 3.

Abstract

α-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid(AMPA)-type glutamate receptors (AMPARs) are the predominant excitatory neurotransmitter receptors in the brain, where they mediate synaptic transmission and plasticity. Excessive AMPAR activation leads to diseases such as epilepsy. AMPAR properties are modulated by auxiliary proteins and foremost by the transmembrane AMPAR regulatory proteins (TARPs). These distribute in unique expression patterns across the brain, rendering AMPAR/TARP complexes promising targets for region-specific therapeutic intervention. TARP γ8 is predominantly expressed in the forebrain and is enriched in the hippocampus, a region associated with temporal lobe epilepsy. Recent high-throughput medicinal chemistry screens have identified multiple promising compounds that selectively target AMPARs associated with γ8 and hold promise for epilepsy treatment. However, how these modulators target the receptor complex is currently unknown. Here, we use a combination of ligand docking, molecular dynamics simulations, and electrophysiology to address this question. We identify a conserved oxindole isostere, shared between three structurally diverse modulators (LY-3130481, JNJ-55511118, and JNJ-61432059) as the major module engaging γ8 by an H-bond to Asn-172 (γ8). The remaining variable region of each molecule likely targets the receptor complex in ligand-selective modes. Functional data reveal parallels in the underlying modulatory action of two prominent compounds. This work will aid development of refined AMPAR epilepsy therapeutics and facilitate to uncover the mechanisms by which TARPs modulate the receptor.

Keywords: AMPA receptor; MD simulations; electrophysiology; glutamate receptor; ionotropic glutamate receptor; synapse; α-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid receptor (AMPA receptor, AMPAR).

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Benzimidazoles / chemistry
  • Benzimidazoles / pharmacology
  • Binding Sites / drug effects
  • Calcium Channels / chemistry
  • Calcium Channels / metabolism*
  • HEK293 Cells
  • Humans
  • Ligands
  • Models, Molecular
  • Molecular Docking Simulation
  • Oxindoles / chemistry*
  • Oxindoles / pharmacology*
  • Protein Binding / drug effects*
  • Protein Interaction Maps / drug effects
  • Rats
  • Receptors, AMPA / chemistry
  • Receptors, AMPA / metabolism*

Substances

  • 5-(2-chloro-6-(trifluoromethoxy)phenyl)-1,3-dihydrobenzimidazol-2-one
  • Benzimidazoles
  • CACNG8 protein, human
  • Calcium Channels
  • Ligands
  • Oxindoles
  • Receptors, AMPA
  • cacng8 protein, rat
  • 2-oxindole

Associated data

  • PDB/6QKZ
  • PDB/6QKC