title: Positive Allosteric Modulators of GluN2A-Containing NMDARs with Distinct Modes of Action and Impacts on Circuit Function.
abstract: To enhance physiological function of NMDA receptors (NMDARs), we identified positive allosteric modulators (PAMs) of NMDARs with selectivity for GluN2A subunit-containing receptors. X-ray crystallography revealed a binding site at the GluN1-GluN2A dimer interface of the extracellular ligand-binding domains (LBDs). Despite the similarity between the LBDs of NMDARs and AMPA receptors (AMPARs), GluN2A PAMs with good selectivity against AMPARs were identified. Potentiation was observed with recombinant triheteromeric GluN1/GluN2A/GluN2B NMDARs and with synaptically activated NMDARs in brain slices from wild-type (WT), but not GluN2A knockout (KO), mice. Individual GluN2A PAMs exhibited variable degrees of glutamate (Glu) dependence, impact on NMDAR Glu EC50, and slowing of channel deactivation. These distinct PAMs also exhibited differential impacts during synaptic plasticity induction. The identification of a new NMDAR modulatory site and characterization of GluN2A-selective PAMs provide powerful molecular tools to dissect NMDAR function and demonstrate the feasibility of a therapeutically desirable type of NMDAR enhancement.
authors.affiliation: Department of Neuroscience, Genentech, Inc., 1 DNA Way, South San Francisco, CA 94080, USA.
authors.forename: David H
authors.initials: DH
authors.lastname: Hackos
authors.affiliation: Department of Structural Biology, Genentech, Inc., 1 DNA Way, South San Francisco, CA 94080, USA.
authors.forename: Patrick J
authors.initials: PJ
authors.lastname: Lupardus
authors.affiliation: Ecole Normale Supérieure, PSL Research University, Institut de Biologie de l'Ecole Normale Supérieure (IBENS), Centre National de la Recherche Scientifique (CNRS) UMR8197, INSERM U1024, 75005 Paris, France.
authors.forename: Teddy
authors.initials: T
authors.lastname: Grand
authors.affiliation: Department of Neuroscience, Genentech, Inc., 1 DNA Way, South San Francisco, CA 94080, USA.
authors.forename: Yelin
authors.initials: Y
authors.lastname: Chen
authors.affiliation: Department of Neuroscience, Genentech, Inc., 1 DNA Way, South San Francisco, CA 94080, USA.
authors.forename: Tzu-Ming
authors.initials: TM
authors.lastname: Wang
authors.affiliation: Department of Biochemical and Cellular Pharmacology, Genentech, Inc., 1 DNA Way, South San Francisco, CA 94080, USA.
authors.forename: Paul
authors.initials: P
authors.lastname: Reynen
authors.affiliation: Department of Biochemical and Cellular Pharmacology, Genentech, Inc., 1 DNA Way, South San Francisco, CA 94080, USA.
authors.forename: Amy
authors.initials: A
authors.lastname: Gustafson
authors.affiliation: Department of Structural Biology, Genentech, Inc., 1 DNA Way, South San Francisco, CA 94080, USA.
authors.forename: Heidi J A
authors.initials: HJ
authors.lastname: Wallweber
authors.affiliation: Department of Discovery Chemistry, Genentech, Inc., 1 DNA Way, South San Francisco, CA 94080, USA.
authors.forename: Matthew
authors.initials: M
authors.lastname: Volgraf
authors.affiliation: Department of Discovery Chemistry, Genentech, Inc., 1 DNA Way, South San Francisco, CA 94080, USA.
authors.forename: Benjamin D
authors.initials: BD
authors.lastname: Sellers
authors.affiliation: Department of Discovery Chemistry, Genentech, Inc., 1 DNA Way, South San Francisco, CA 94080, USA.
authors.forename: Jacob B
authors.initials: JB
authors.lastname: Schwarz
authors.affiliation: Ecole Normale Supérieure, PSL Research University, Institut de Biologie de l'Ecole Normale Supérieure (IBENS), Centre National de la Recherche Scientifique (CNRS) UMR8197, INSERM U1024, 75005 Paris, France.
authors.forename: Pierre
authors.initials: P
authors.lastname: Paoletti
authors.affiliation: Department of Neuroscience, Genentech, Inc., 1 DNA Way, South San Francisco, CA 94080, USA.
authors.forename: Morgan
authors.initials: M
authors.lastname: Sheng
authors.affiliation: Department of Neuroscience, Genentech, Inc., 1 DNA Way, South San Francisco, CA 94080, USA.
authors.forename: Qiang
authors.initials: Q
authors.lastname: Zhou
authors.affiliation: Department of Neuroscience, Genentech, Inc., 1 DNA Way, South San Francisco, CA 94080, USA. Electronic address: hanson.jesse@gene.com.
authors.forename: Jesse E
authors.initials: JE
authors.lastname: Hanson
mesh_terms: D000494:Allosteric Regulation; D000818:Animals; D001665:Binding Sites / Q000235:genetics; D016466:CHO Cells; D002118:Calcium / Q000378:metabolism; D003412:Cricetulus; D018360:Crystallography, X-Ray; D018683:Excitatory Amino Acid Agents / Q000494:pharmacology; D019706:Excitatory Postsynaptic Potentials / Q000187:drug effects / Q000235:genetics; D057809:HEK293 Cells; D006624:Hippocampus / Q000166:cytology; D006801:Humans; D008564:Membrane Potentials / Q000187:drug effects / Q000235:genetics; D051379:Mice; D008810:Mice, Inbred C57BL; D008822:Mice, Transgenic; D008958:Models, Molecular; D009415:Nerve Net / Q000502:physiology*; D009474:Neurons / Q000187:drug effects / Q000502:physiology*; D016194:Receptors, N-Methyl-D-Aspartate / Q000235:genetics / Q000378:metabolism*
chemical_list: D002118:Calcium; D018683:Excitatory Amino Acid Agents; D016194:Receptors, N-Methyl-D-Aspartate