Structural Determinants and Mechanism of Action of a GluN2C-selective NMDA Receptor Positive Allosteric Modulator

Khatri, A., Burger, P. B., Swanger, S. A., Hansen, K. B., Zimmerman, S., Karakas, E., Liotta, D. C., Furukawa, H., Snyder, J. P., Traynelis, S. F. (2014) Structural Determinants and Mechanism of Action of a GluN2C-selective NMDA Receptor Positive Allosteric Modulator. Mol Pharmacol, 86 (5). pp. 548-560. ISSN 0026-895x

URL: http://www.ncbi.nlm.nih.gov/pubmed/25205677
DOI: 10.1124/mol.114.094516

Abstract

NMDA receptors are tetrameric complexes of GluN1, GluN2A-D, and GluN3A-B subunits and are involved in normal brain function and neurological disorders. We have identified a novel class of stereo-selective pyrrolidinone (PYD) positive allosteric modulators for GluN2C-containing NMDA receptors, exemplified by methyl 4-(3-acetyl-4-hydroxy-1-(2-(2-methyl-1H-indol-3-yl)ethyl)-5-oxo-2,5-dihydro-1H-py rrol-2-yl)benzoate. Here we explore the site and mechanism of action of a prototypical analogue PYD-106, which at 30 muM does not alter responses of NMDA receptors containing GluN2A, GluN2B, and GluN2D, and has no effect on AMPA and kainate receptors. Co-application of 50 muM PYD-106 with a maximally effective concentration of glutamate and glycine increases the response of GluN1/GluN2C NMDA receptors in HEK-293 cells to 221% of that obtained in the absence of PYD (taken as 100%). Evaluation of the concentration-dependence of this enhancement revealed an EC50 value for PYD of 13 muM. PYD-106 increased opening frequency and open time of single channel currents activated by maximally effective concentrations of agonist, but only had modest effects on glutamate and glycine EC50. PYD-106 selectively enhanced the responses of diheteromeric GluN1/GluN2C receptors, but not triheteromeric GluN1/GluN2A/GluN2C receptors. Inclusion of residues encoded by GluN1-exon5 attenuated the effects of PYD. Three GluN2C residues (Arg194, Ser470, Lys470), at which mutagenesis virtually eliminated PYD function, line a cavity at the interface of the ligand binding and the amino terminal domains in a homology model of GluN1/GluN2C built from crystallographic data on GluN1/GluN2B. We propose that this domain interface constitutes a new allosteric modulatory site on the NMDA receptor.

Item Type: Paper
Subjects: bioinformatics > genomics and proteomics > genetics & nucleic acid processing > protein structure, function, modification > protein types > NMDA receptor
structural biology
CSHL Authors:
Communities: CSHL labs > Furukawa lab
Depositing User: Matt Covey
Date Deposited: 24 Sep 2014 14:33
Last Modified: 30 Aug 2017 14:45
PMCID: PMC4201136
Related URLs:
URI: http://repository.cshl.edu/id/eprint/30813

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