Two N-glycosylation Sites in the GluN1 Subunit Are Essential for Releasing N-methyl-d-aspartate (NMDA) Receptors from the Endoplasmic Reticulum

• Published in: The Journal of biological chemistry 2015-07, Vol.290 (30), p.18379-18390
• Main Authors: Lichnerova, Katarina, Kaniakova, Martina, Park, Seung Pyo, Skrenkova, Kristyna, Wang, Ya-Xian, Petralia, Ronald S., Suh, Young Ho, Horak, Martin
• Format: Article
• Language: English
• Subjects: Animals; Chlorocebus aethiops; COS Cells; endoplasmic reticulum (ER); Endoplasmic Reticulum - metabolism; glutamate receptor; Glycosylation; Golgi Apparatus - metabolism; HEK293 Cells; Humans; N-Methylaspartate - chemistry; N-Methylaspartate - metabolism; Neurobiology; neuron; Neurons - chemistry; Neurons - metabolism; Polysaccharides - metabolism; Rats; Receptors, N-Methyl-D-Aspartate - chemistry; Receptors, N-Methyl-D-Aspartate - metabolism; synapse; Synapses - metabolism; Synaptic Transmission; trafficking
• ISSN: 0021-9258; 1083-351X
• DOI: 10.1074/jbc.M115.656546

NMDA receptors (NMDARs) comprise a subclass of neurotransmitter receptors whose surface expression is regulated at multiple levels, including processing in the endoplasmic reticulum (ER), intracellular trafficking via the Golgi apparatus, internalization, recycling, and degradation. With respect to early processing, NMDARs are regulated by the availability of GluN subunits within the ER, the presence of ER retention and export signals, and posttranslational modifications, including phosphorylation and palmitoylation. However, the role of N-glycosylation, one of the most common posttranslational modifications, in regulating NMDAR processing has not been studied in detail. Using biochemistry, confocal and electron microscopy, and electrophysiology in conjunction with a lentivirus-based molecular replacement strategy, we found that NMDARs are released from the ER only when two asparagine residues in the GluN1 subunit (Asn-203 and Asn-368) are N-glycosylated. Although the GluN2A and GluN2B subunits are also N-glycosylated, their N-glycosylation sites do not appear to be essential for surface delivery of NMDARs. Furthermore, we found that removing N-glycans from native NMDARs altered the receptor affinity for glutamate. Our results suggest a novel mechanism by which neurons ensure that postsynaptic membranes contain sufficient numbers of functional NMDARs. Regulation of NMDA receptors is critical for excitatory neurotransmission. N-glycans are essential for NMDA receptor release from the endoplasmic reticulum and for receptor affinity for the agonist. N-glycosylation regulates the trafficking and function of NMDA receptors. We identified a novel mechanism that could ensure that postsynaptic membranes contain sufficient numbers of NMDA receptors.