Shining Light on an mGlu5 Photoswitchable NAM: A Theoretical Perspective

Curr Neuropharmacol. 2016;14(5):441-54. doi: 10.2174/1570159x13666150407231417.

Abstract

Metabotropic glutamate receptors (mGluRs) are important drug targets because of their involvement in several neurological diseases. Among mGluRs, mGlu5 is a particularly high-profile target because its positive or negative allosteric modulation can potentially treat schizophrenia or anxiety and chronic pain, respectively. Here, we computationally and experimentally probe the functional binding of a novel photoswitchable mGlu5 NAM, termed alloswitch-1, which loses its NAM functionality under violet light. We show alloswitch-1 binds deep in the allosteric pocket in a similar fashion to mavoglurant, the co-crystallized NAM in the mGlu5 transmembrane domain crystal structure. Alloswitch-1, like NAM 2-Methyl-6-(phenylethynyl)pyridine (MPEP), is significantly affected by P655M mutation deep in the allosteric pocket, eradicating its functionality. In MD simulations, we show alloswitch-1 and MPEP stabilize the co-crystallized water molecule located at the bottom of the allosteric site that is seemingly characteristic of the inactive receptor state. Furthermore, both NAMs form H-bonds with S809 on helix 7, which may constitute an important stabilizing interaction for NAM-induced mGlu5 inactivation. Alloswitch-1, through isomerization of its amide group from trans to cis is able to form an additional interaction with N747 on helix 5. This may be an important interaction for amide-containing mGlu5 NAMs, helping to stabilize their binding in a potentially unusual cis-amide state. Simulated conformational switching of alloswitch-1 in silico suggests photoisomerization of its azo group from trans to cis may be possible within the allosteric pocket. However, photoexcited alloswitch-1 binds in an unstable fashion, breaking H-bonds with the protein and destabilizing the co-crystallized water molecule. This suggests photoswitching may have destabilizing effects on mGlu5 binding and functionality.

Publication types

  • Validation Study

MeSH terms

  • Allosteric Regulation*
  • Allosteric Site
  • Excitatory Amino Acid Antagonists / pharmacology
  • HEK293 Cells
  • Humans
  • Hydrogen Bonding
  • Isomerism
  • Light*
  • Molecular Docking Simulation
  • Molecular Dynamics Simulation
  • Mutation
  • Photochemical Processes*
  • Protein Binding
  • Protein Conformation
  • Protein Stability
  • Pyridines / pharmacology
  • Receptor, Metabotropic Glutamate 5 / antagonists & inhibitors
  • Receptor, Metabotropic Glutamate 5 / genetics
  • Receptor, Metabotropic Glutamate 5 / metabolism*
  • Receptor, Metabotropic Glutamate 5 / radiation effects*
  • Water / chemistry

Substances

  • Excitatory Amino Acid Antagonists
  • Pyridines
  • Receptor, Metabotropic Glutamate 5
  • Water
  • 6-methyl-2-(phenylethynyl)pyridine