NMDA receptors are selectively partitioned into complexes and supercomplexes during synapse maturation.
How neuronal proteomes self-organize is poorly understood because of their inherent molecular and cellular complexity. Here, focusing on mammalian synapses we use blue-native PAGE and 'gene-tagging' of GluN1 to report the first biochemical purification of endogenous NMDA receptors (NMDARs) directly from adult mouse brain. We show that NMDARs partition between two discrete populations of receptor complexes and ∼1.5 MDa supercomplexes. We tested the assembly mechanism with six mouse mutants, which indicates a tripartite requirement of GluN2B, PSD93 and PSD95 gate the incorporation of receptors into ∼1.5 MDa supercomplexes, independent of either canonical PDZ-ligands or GluN2A. Supporting the essential role of GluN2B, quantitative gene-tagging revealed a fourfold molar excess of GluN2B over GluN2A in adult forebrain. NMDAR supercomplexes are assembled late in postnatal development and triggered by synapse maturation involving epigenetic and activity-dependent mechanisms. Finally, screening the quaternary organization of 60 native proteins identified numerous discrete supercomplexes that populate the mammalian synapse.
Centre for Clinical Brain Sciences, University of Edinburgh, Chancellor's Building, 49 Little France Crescent, Edinburgh EH16 4SB, UK.|MRC Laboratory of Molecular Biology, Hills Road, Cambridge CB2 0QH, UK.
René A W
RA
Frank
Centre for Clinical Brain Sciences, University of Edinburgh, Chancellor's Building, 49 Little France Crescent, Edinburgh EH16 4SB, UK.
Noboru H
NH
Komiyama
RIKEN-MIT Center for Neural Circuit Genetics at the Picower Institute for Learning and Memory, Department of Biology, Massachusetts Institute of Technology, Cambridge, Massachusetts 02139, USA.|Howard Hughes Medical Institute, Massachusetts Institute of Technology, Cambridge, Massachusetts 02139, USA.
Tomás J
TJ
Ryan
Centre for Clinical Brain Sciences, University of Edinburgh, Chancellor's Building, 49 Little France Crescent, Edinburgh EH16 4SB, UK.
Fei
F
Zhu
Department of Physiology, David Geffen School of Medicine and UCLA Integrative Center for Learning and Memory, University of California Los Angeles, Los Angeles, California 90095, USA.
Thomas J
TJ
O'Dell
Centre for Clinical Brain Sciences, University of Edinburgh, Chancellor's Building, 49 Little France Crescent, Edinburgh EH16 4SB, UK.
Seth G N
SG
Grant
D000818:Animals; D002478:Cells, Cultured; D057809:HEK293 Cells; D006801:Humans; D008822:Mice, Transgenic; D046912:Multiprotein Complexes / Q000235:genetics / Q000378:metabolism*; D009154:Mutation; D009419:Nerve Tissue Proteins / Q000235:genetics / Q000378:metabolism; D016548:Prosencephalon / Q000166:cytology / Q000196:embryology / Q000378:metabolism; D011485:Protein Binding; D020543:Proteome / Q000378:metabolism; D040901:Proteomics; D016194:Receptors, N-Methyl-D-Aspartate / Q000235:genetics / Q000378:metabolism*; D013569:Synapses / Q000235:genetics / Q000378:metabolism*
D046912:Multiprotein Complexes; D009419:Nerve Tissue Proteins; D020543:Proteome; D016194:Receptors, N-Methyl-D-Aspartate; C121673:Gprin1 protein, mouse; C121001:NR2B NMDA receptor