The NMDA receptor intracellular C-terminal domains reciprocally interact with allosteric modulators

Kiran Sapkota, Kim Dore, Kang Tang, Mark Irvine, Guangyu Fang, Erica S. Burnell, Roberto Malinow, David E. Jane, Daniel T. Monaghan

Research output: Contribution to journalArticle

Abstract

N-methyl-D-aspartate receptors (NMDARs) have multiple prominent roles in CNS function but their excessive or insufficient activity contributes to neuropathological/psychiatric disorders. Consequently, a variety of positive and negative allosteric modulators (PAMs and NAMs, respectively) have recently been developed. Although these modulators bind to extracellular domains, in the present report we find that the NMDAR's intracellular C-terminal domains (CTDs) significantly influence PAM/NAM activity. GluN2 CTD deletion robustly affected NAM and PAM activity with both enhancing and inhibiting effects that were compound-specific and NMDAR subunit-specific. In three cases, individual PAMs became NAMs at specific GluN2-truncated receptors. In contrast to GluN2, GluN1 CTD removal only reduced PAM activity of UBP684 and CIQ, and did not affect NAM activity. Consistent with these findings, agents altering phosphorylation state or intracellular calcium levels displayed receptor-specific and compound-specific effects on PAM activity. It is possible that the GluN2′s M4 domain transmits intracellular modulatory signals from the CTD to the M1/M4 channel gating machinery and that this site is a point of convergence in the direct or indirect actions of several PAMs/NAMs thus rendering them sensitive to CTD status. Thus, allosteric modulators are likely to have a marked and varied sensitivity to post-translational modifications, protein-protein associations, and intracellular ions. The interaction between PAM activity and NMDAR CTDs appears reciprocal. GluN1 CTD-deletion eliminated UBP684, but not pregnenolone sulfate (PS), PAM activity. And, in the absence of agonists, UBP684, but not PS, was able to promote movement of fluorescently-tagged GluN1-CTDs. Thus, it may be possible to pharmacologically target NMDAR metabotropic activity in the absence of channel activation.

Original languageEnglish (US)
Pages (from-to)140-153
Number of pages14
JournalBiochemical Pharmacology
Volume159
DOIs
StatePublished - Jan 2019

Fingerprint

Pulse amplitude modulation
N-Methyl-D-Aspartate Receptors
Modulators
Post Translational Protein Processing
Psychiatry
Phosphorylation
Ions
Calcium
Proteins
Machinery
Chemical activation
pregnenolone sulfate

Keywords

  • Allosteric modulators
  • C-terminal domain
  • Desensitization
  • Fluorescence resonance energy transfer
  • N-methyl-D-aspartate
  • Phosphorylation

ASJC Scopus subject areas

  • Biochemistry
  • Pharmacology

Cite this

The NMDA receptor intracellular C-terminal domains reciprocally interact with allosteric modulators. / Sapkota, Kiran; Dore, Kim; Tang, Kang; Irvine, Mark; Fang, Guangyu; Burnell, Erica S.; Malinow, Roberto; Jane, David E.; Monaghan, Daniel T.

In: Biochemical Pharmacology, Vol. 159, 01.2019, p. 140-153.

Research output: Contribution to journalArticle

Sapkota, Kiran ; Dore, Kim ; Tang, Kang ; Irvine, Mark ; Fang, Guangyu ; Burnell, Erica S. ; Malinow, Roberto ; Jane, David E. ; Monaghan, Daniel T. / The NMDA receptor intracellular C-terminal domains reciprocally interact with allosteric modulators. In: Biochemical Pharmacology. 2019 ; Vol. 159. pp. 140-153.
@article{4123d05c3ec94d479c65c2fee9bfdcab,
title = "The NMDA receptor intracellular C-terminal domains reciprocally interact with allosteric modulators",
abstract = "N-methyl-D-aspartate receptors (NMDARs) have multiple prominent roles in CNS function but their excessive or insufficient activity contributes to neuropathological/psychiatric disorders. Consequently, a variety of positive and negative allosteric modulators (PAMs and NAMs, respectively) have recently been developed. Although these modulators bind to extracellular domains, in the present report we find that the NMDAR's intracellular C-terminal domains (CTDs) significantly influence PAM/NAM activity. GluN2 CTD deletion robustly affected NAM and PAM activity with both enhancing and inhibiting effects that were compound-specific and NMDAR subunit-specific. In three cases, individual PAMs became NAMs at specific GluN2-truncated receptors. In contrast to GluN2, GluN1 CTD removal only reduced PAM activity of UBP684 and CIQ, and did not affect NAM activity. Consistent with these findings, agents altering phosphorylation state or intracellular calcium levels displayed receptor-specific and compound-specific effects on PAM activity. It is possible that the GluN2′s M4 domain transmits intracellular modulatory signals from the CTD to the M1/M4 channel gating machinery and that this site is a point of convergence in the direct or indirect actions of several PAMs/NAMs thus rendering them sensitive to CTD status. Thus, allosteric modulators are likely to have a marked and varied sensitivity to post-translational modifications, protein-protein associations, and intracellular ions. The interaction between PAM activity and NMDAR CTDs appears reciprocal. GluN1 CTD-deletion eliminated UBP684, but not pregnenolone sulfate (PS), PAM activity. And, in the absence of agonists, UBP684, but not PS, was able to promote movement of fluorescently-tagged GluN1-CTDs. Thus, it may be possible to pharmacologically target NMDAR metabotropic activity in the absence of channel activation.",
keywords = "Allosteric modulators, C-terminal domain, Desensitization, Fluorescence resonance energy transfer, N-methyl-D-aspartate, Phosphorylation",
author = "Kiran Sapkota and Kim Dore and Kang Tang and Mark Irvine and Guangyu Fang and Burnell, {Erica S.} and Roberto Malinow and Jane, {David E.} and Monaghan, {Daniel T.}",
year = "2019",
month = "1",
doi = "10.1016/j.bcp.2018.11.018",
language = "English (US)",
volume = "159",
pages = "140--153",
journal = "Biochemical Pharmacology",
issn = "0006-2952",
publisher = "Elsevier Inc.",

}

TY - JOUR

T1 - The NMDA receptor intracellular C-terminal domains reciprocally interact with allosteric modulators

AU - Sapkota, Kiran

AU - Dore, Kim

AU - Tang, Kang

AU - Irvine, Mark

AU - Fang, Guangyu

AU - Burnell, Erica S.

AU - Malinow, Roberto

AU - Jane, David E.

AU - Monaghan, Daniel T.

PY - 2019/1

Y1 - 2019/1

N2 - N-methyl-D-aspartate receptors (NMDARs) have multiple prominent roles in CNS function but their excessive or insufficient activity contributes to neuropathological/psychiatric disorders. Consequently, a variety of positive and negative allosteric modulators (PAMs and NAMs, respectively) have recently been developed. Although these modulators bind to extracellular domains, in the present report we find that the NMDAR's intracellular C-terminal domains (CTDs) significantly influence PAM/NAM activity. GluN2 CTD deletion robustly affected NAM and PAM activity with both enhancing and inhibiting effects that were compound-specific and NMDAR subunit-specific. In three cases, individual PAMs became NAMs at specific GluN2-truncated receptors. In contrast to GluN2, GluN1 CTD removal only reduced PAM activity of UBP684 and CIQ, and did not affect NAM activity. Consistent with these findings, agents altering phosphorylation state or intracellular calcium levels displayed receptor-specific and compound-specific effects on PAM activity. It is possible that the GluN2′s M4 domain transmits intracellular modulatory signals from the CTD to the M1/M4 channel gating machinery and that this site is a point of convergence in the direct or indirect actions of several PAMs/NAMs thus rendering them sensitive to CTD status. Thus, allosteric modulators are likely to have a marked and varied sensitivity to post-translational modifications, protein-protein associations, and intracellular ions. The interaction between PAM activity and NMDAR CTDs appears reciprocal. GluN1 CTD-deletion eliminated UBP684, but not pregnenolone sulfate (PS), PAM activity. And, in the absence of agonists, UBP684, but not PS, was able to promote movement of fluorescently-tagged GluN1-CTDs. Thus, it may be possible to pharmacologically target NMDAR metabotropic activity in the absence of channel activation.

AB - N-methyl-D-aspartate receptors (NMDARs) have multiple prominent roles in CNS function but their excessive or insufficient activity contributes to neuropathological/psychiatric disorders. Consequently, a variety of positive and negative allosteric modulators (PAMs and NAMs, respectively) have recently been developed. Although these modulators bind to extracellular domains, in the present report we find that the NMDAR's intracellular C-terminal domains (CTDs) significantly influence PAM/NAM activity. GluN2 CTD deletion robustly affected NAM and PAM activity with both enhancing and inhibiting effects that were compound-specific and NMDAR subunit-specific. In three cases, individual PAMs became NAMs at specific GluN2-truncated receptors. In contrast to GluN2, GluN1 CTD removal only reduced PAM activity of UBP684 and CIQ, and did not affect NAM activity. Consistent with these findings, agents altering phosphorylation state or intracellular calcium levels displayed receptor-specific and compound-specific effects on PAM activity. It is possible that the GluN2′s M4 domain transmits intracellular modulatory signals from the CTD to the M1/M4 channel gating machinery and that this site is a point of convergence in the direct or indirect actions of several PAMs/NAMs thus rendering them sensitive to CTD status. Thus, allosteric modulators are likely to have a marked and varied sensitivity to post-translational modifications, protein-protein associations, and intracellular ions. The interaction between PAM activity and NMDAR CTDs appears reciprocal. GluN1 CTD-deletion eliminated UBP684, but not pregnenolone sulfate (PS), PAM activity. And, in the absence of agonists, UBP684, but not PS, was able to promote movement of fluorescently-tagged GluN1-CTDs. Thus, it may be possible to pharmacologically target NMDAR metabotropic activity in the absence of channel activation.

KW - Allosteric modulators

KW - C-terminal domain

KW - Desensitization

KW - Fluorescence resonance energy transfer

KW - N-methyl-D-aspartate

KW - Phosphorylation

UR - http://www.scopus.com/inward/record.url?scp=85058054700&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=85058054700&partnerID=8YFLogxK

U2 - 10.1016/j.bcp.2018.11.018

DO - 10.1016/j.bcp.2018.11.018

M3 - Article

C2 - 30503374

AN - SCOPUS:85058054700

VL - 159

SP - 140

EP - 153

JO - Biochemical Pharmacology

JF - Biochemical Pharmacology

SN - 0006-2952

ER -