Subunit arrangement and phenylethanolamine binding in GluN1/GluN2B NMDA receptors

Karakaş, Erkan; Simorowski, Noriko; Furukawa, Hiro

doi:10.1038/nature10180

Cited by 302 publications

(453 citation statements)

References 27 publications

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“…To genetically encode photoreactive UAAs, we chose the GluN1-Y109 site, which is situated at the NTD upper lobe-upper lobe (UL/UL) dimer interface according to the X-ray crystal structure of the GluN1/ GluN2B NTD dimer (16) (Fig. 1A).…”

Section: Resultsmentioning

confidence: 99%

“…We first tested sensitivity to ifenprodil, the prototypical GluN2B-selective inhibitor, which binds the dimer interface between GluN1 and GluN2B NTDs (16). Ifenprodil sensitivity of photo-treated AzF-encoded receptors was markedly reduced compared with the situation before UV treatment (∼10-fold increase in IC 50 ) (SI Appendix, Fig.…”

Section: Resultsmentioning

confidence: 99%

“…Besides having essential functions in receptor assembly (10,11), recent studies of the NTDs have also revealed that the individual GluN2 (12)(13)(14) and GluN1 (15) NTDs fine-tune NMDAR gating and pharmacological properties by undergoing large-range conformational changes. The recent X-ray crystal structure of a GluN1/GluN2B NTD complex reveals a unique arrangement of the two NTD protomers with intersubunit interactions distinct from those observed in AMPA and kainate receptors (16,17). However, the importance of these dimer interfaces in the subunit-specific receptor regulation is poorly understood.…”

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confidence: 99%

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Genetically encoding a light switch in an ionotropic glutamate receptor reveals subunit-specific interfaces

Zhu

Riou

Yao

et al. 2014

Proc. Natl. Acad. Sci. U.S.A.

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Reprogramming receptors to artificially respond to light has strong potential for molecular studies and interrogation of biological functions. Here, we design a light-controlled ionotropic glutamate receptor by genetically encoding a photoreactive unnatural amino acid (UAA). The photo-cross-linker p-azido-L-phenylalanine (AzF) was encoded in NMDA receptors (NMDARs), a class of glutamate-gated ion channels that play key roles in neuronal development and plasticity. AzF incorporation in the obligatory GluN1 subunit at the GluN1/GluN2B N-terminal domain (NTD) upper lobe dimer interface leads to an irreversible allosteric inhibition of channel activity upon UV illumination. In contrast, when pairing the UAA-containing GluN1 subunit with the GluN2A subunit, light-dependent inactivation is completely absent. By combining electrophysiological and biochemical analyses, we identify subunit-specific structural determinants at the GluN1/GluN2 NTD dimer interfaces that critically dictate UV-controlled inactivation. Our work reveals that the two major NMDAR subtypes differ in their ectodomain-subunit interactions, in particular their electrostatic contacts, resulting in GluN1 NTD coupling more tightly to the GluN2B NTD than to the GluN2A NTD. It also paves the way for engineering light-sensitive ligand-gated ion channels with subtype specificity through the genetic code expansion.neurotransmitter receptor | protein structure-function I ntroducing light-sensitive moieties into proteins provides a powerful approach to investigate molecular mechanisms as well as biological functions with high temporal and spatial resolution (1, 2). An attractive strategy to engineer light responsiveness relies on the use of photoreactive unnatural amino acids (UAAs), allowing site-specific incorporation in a protein target. The methodology relies on the read-through of an unassigned codon (commonly the amber stop codon) in an mRNA by a suppressor tRNA aminoacylated with a desired UAA. Using this approach, UAAs with unique chemical functionalities including light-sensitivity have been successfully incorporated into ion channels and neurotransmitter receptors, significantly contributing to our understanding of receptor function (3, 4). However, the challenging synthesis of the chemically acylated tRNA has limited the general applicability of the approach. The recent development of genetically engineered suppressor tRNA/ aminoacyl-tRNA synthetase pairs with altered amino acid specificity allowed for aminoacylation in the expression system in situ. This method provided a major step forward by advancing the UAA technology to the all-genetic-based level, also known as "the genetic-code expansion" (5-7).Here, we present the design of a light-sensitive ionotropic glutamate receptor (iGluR) through the genetic incorporation of a photoreactive UAA. Our approach takes advantage of the recent development of the genetic-code expansion in Xenopus oocytes (8), which is a classical vehicle for heterologous expression and functional characterization of ligand-g...

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Section: Resultsmentioning

confidence: 99%

Section: Resultsmentioning

confidence: 99%

mentioning

confidence: 99%

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Genetically encoding a light switch in an ionotropic glutamate receptor reveals subunit-specific interfaces

Zhu

Riou

Yao

et al. 2014

Proc. Natl. Acad. Sci. U.S.A.

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“…49 The simulation was performed according to the SCARE induced fit docking protocol that has been thoroughly described by Bottegoni and colleagues, 50 and it is only briefly summarized here. SCARE implements the flexibility of the receptor in the simulation generating different local versions of the binding pocket.…”

Section: Journal Of Medicinal Chemistrymentioning

confidence: 99%

Combining Galantamine and Memantine in Multitargeted, New Chemical Entities Potentially Useful in Alzheimer’s Disease

et al. 2012

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Herein we report on a novel series of multitargeted compounds obtained by linking together galantamine and memantine. The compounds were designed by taking advantage of the crystal structures of acetylcholinesterase (AChE) in complex with galantamine derivatives. Sixteen novel derivatives were synthesized, using spacers of different lengths and chemical composition. The molecules were then tested as inhibitors of AChE and as binders of the N-methyl-D-aspartate (NMDA) receptor (NMDAR). Some of the new compounds were nanomolar inhibitors of AChE and showed micromolar affinities for NMDAR. All compounds were also tested for selectivity toward NMDAR containing the 2B subunit (NR2B). Some of the new derivatives showed a micromolar affinity for NR2B. Finally, selected compounds were tested using a cellbased assay to measure their neuroprotective activity. Three of them showed a remarkable neuroprotective profile, inhibiting the NMDAinduced neurotoxicity at subnanomolar concentrations (e.g., 5, named memagal, IC 50 = 0.28 nM).

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“…A negative allosteric modulator (NAM) of the NR2B subtype of NMDA receptor, CP-101,606 Mott et al, 1998), also evidenced an antidepressant response in patients (Preskorn et al, 2008). Compounds of the NR2B NAM class bind at the interface of the NR2B/ NR1 amino terminal domains to allosterically reduce channel-opening probability to inhibit ion flux and functionally inhibit receptor activity (Karakas et al, 2011;Traynelis et al, 2010). Like ketamine, the response to CP-101,606 developed rapidly after a short drug infusion and was sustained well after the drug was cleared from the body.…”

Section: Introductionmentioning

confidence: 99%

Differential Effects of an NR2B NAM and Ketamine on Synaptic Potentiation and Gamma Synchrony: Relevance to Rapid-Onset Antidepressant Efficacy

Nagy

Stoiljković

Menniti

et al. 2015

Neuropsychopharmacol

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Ketamine, a pan-NMDA receptor channel blocker, and CP-101,606, an NR2B-selective negative allosteric modulator, have antidepressant effects in humans that develop rapidly after the drugs are cleared from the body. It has been proposed that the antidepressant effect of ketamine results from delayed synaptic potentiation. To further investigate this hypothesis and potential mechanistic underpinnings we compared the effects of ketamine and CP-101,606 on neurophysiological biomarkers in rats immediately after drug administration and after the drugs had been eliminated. Local field and auditory-evoked potentials (AEPs) were recorded from primary auditory cortex and hippocampus in freely moving rats. Effects of different doses of ketamine or CP-101,606 were evaluated on amplitude of AEPs, auditory gating, and absolute power of delta and gamma oscillations 5-30 min (drug-on) and 5-6 h (drug-off) after systemic administration. Both ketamine and CP-101,606 significantly enhanced AEPs in cortex and hippocampus in the drug-off phase. In contrast, ketamine but not CP-101,606 disrupted auditory gating and increased gamma-band power during the drug-on period. Although both drugs affected delta power, these changes did not correlate with increase in AEPs in the drug-off phase. Our findings show that both ketamine and CP-101,606 augment AEPs after drug elimination, consistent with synaptic potentiation as a mechanism for antidepressant efficacy. However, these drugs had different acute effects on neurophysiological parameters. These results have implications for understanding the underlying mechanisms for the rapid-onset antidepressant effects of NMDA receptor inhibition and for the use of electrophysiological measures as translatable biomarkers.

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Subunit arrangement and phenylethanolamine binding in GluN1/GluN2B NMDA receptors

Cited by 302 publications

References 27 publications

Genetically encoding a light switch in an ionotropic glutamate receptor reveals subunit-specific interfaces

Genetically encoding a light switch in an ionotropic glutamate receptor reveals subunit-specific interfaces

Combining Galantamine and Memantine in Multitargeted, New Chemical Entities Potentially Useful in Alzheimer’s Disease

Differential Effects of an NR2B NAM and Ketamine on Synaptic Potentiation and Gamma Synchrony: Relevance to Rapid-Onset Antidepressant Efficacy

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