Influence of a threonine residue in the S2 ligand binding domain in determining agonist potency and deactivation rate of recombinant NR1a/NR2D NMDA receptors. / Chen, P. E.; Johnston, A. R.; Mok, M. H.; Schoepfer, R.; Wyllie, D. J.

In: Journal of Physiology, Vol. 558, No. Pt 1, 2004, p. 45-58.

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Influence of a threonine residue in the S2 ligand binding domain in determining agonist potency and deactivation rate of recombinant NR1a/NR2D NMDA receptors. / Chen, P. E.; Johnston, A. R.; Mok, M. H.; Schoepfer, R.; Wyllie, D. J.

In: Journal of Physiology, Vol. 558, No. Pt 1, 2004, p. 45-58.

Research output: Contribution to journalArticlepeer-review

Harvard

Chen, PE, Johnston, AR, Mok, MH, Schoepfer, R & Wyllie, DJ 2004, 'Influence of a threonine residue in the S2 ligand binding domain in determining agonist potency and deactivation rate of recombinant NR1a/NR2D NMDA receptors', Journal of Physiology, vol. 558, no. Pt 1, pp. 45-58.

APA

Chen, P. E., Johnston, A. R., Mok, M. H., Schoepfer, R., & Wyllie, D. J. (2004). Influence of a threonine residue in the S2 ligand binding domain in determining agonist potency and deactivation rate of recombinant NR1a/NR2D NMDA receptors. Journal of Physiology, 558(Pt 1), 45-58.

Vancouver

Author

Chen, P. E. ; Johnston, A. R. ; Mok, M. H. ; Schoepfer, R. ; Wyllie, D. J. / Influence of a threonine residue in the S2 ligand binding domain in determining agonist potency and deactivation rate of recombinant NR1a/NR2D NMDA receptors. In: Journal of Physiology. 2004 ; Vol. 558, No. Pt 1. pp. 45-58.

BibTeX

@article{fa9019a031c74bfb9fd337e7c2869af7,
title = "Influence of a threonine residue in the S2 ligand binding domain in determining agonist potency and deactivation rate of recombinant NR1a/NR2D NMDA receptors",
abstract = "NR1/NR2D NMDA receptors display unusually slow deactivation kinetics which may be critical for their role as extrasynaptic receptors. A threonine to alanine point mutation has been inserted at amino acid position 692 of the NR2D subunit (T692A). Recombinant NR1a/NR2D(T692A) NMDA receptors have been expressed in Xenopus laevis oocytes and their pharmacological and single-channel properties examined using two-electrode voltage-clamp and patch-clamp recording techniques. Glutamate dose-response curves from NR1a/NR2D(T692A) receptor channels produced an approximately 1600-fold reduction in glutamate potency compared to wild-type NR1a/NR2D receptors. There was no change in Hill slopes or gross reduction in mean maximal currents recorded in oocytes expressing either wild-type or mutant receptors. The mutation did not affect the potency of the co-agonist glycine. The shifts in potency produced by NR2D(T692A) containing receptors when activated by other glutamate-site agonists such as aspartate or NMDA were 30- to 60-fold compared to wild-type. Single-channel conductance levels of NR1a/NR2D(T692A) mutant receptors were indistinguishable from wild-type NR2D-containing channels. Additionally NR1a/NR2D(T692A) receptors showed the transitional asymmetry that is characteristic of NR2D-containing NMDA receptors. Rapid applications of glutamate on outside-out patches containing NR1a/NR2D(T692A) receptors produced macroscopic current deactivations that were about 60-fold faster than wild-type NR1a/NR2D receptors. Our results suggest that this conserved threonine residue plays a crucial role in ligand binding to NMDA NR2 receptor subunits and supports the idea that the slow decay kinetics associated with NR1a/NR2D NMDA receptors can be explained by the slow dissociation of glutamate from this NMDA receptor subtype.",
keywords = "Animals Binding Sites/physiology Glutamic Acid/pharmacology Ion Channel Gating/drug effects/physiology Ligands Mutagenesis, Site-Directed Oocytes/physiology Patch-Clamp Techniques Protein Structure, Tertiary Rats Receptors, N-Methyl-D-Aspartate/agonists/*genetics/*metabolism Threonine/genetics Xenopus laevis",
author = "Chen, {P. E.} and Johnston, {A. R.} and Mok, {M. H.} and R. Schoepfer and Wyllie, {D. J.}",
note = "Chen, Philip E Johnston, Alexander R Mok, M H Selina Schoepfer, Ralf Wyllie, David J A Research Support, Non-U.S. Gov't England The Journal of physiology J Physiol. 2004 Jul 1;558(Pt 1):45-58. Epub 2004 Apr 23.",
year = "2004",
language = "English",
volume = "558",
pages = "45--58",
journal = "Journal of Physiology ",
issn = "0022-3751",
publisher = "Wiley-Blackwell",
number = "Pt 1",

}

RIS

TY - JOUR

T1 - Influence of a threonine residue in the S2 ligand binding domain in determining agonist potency and deactivation rate of recombinant NR1a/NR2D NMDA receptors

AU - Chen, P. E.

AU - Johnston, A. R.

AU - Mok, M. H.

AU - Schoepfer, R.

AU - Wyllie, D. J.

N1 - Chen, Philip E Johnston, Alexander R Mok, M H Selina Schoepfer, Ralf Wyllie, David J A Research Support, Non-U.S. Gov't England The Journal of physiology J Physiol. 2004 Jul 1;558(Pt 1):45-58. Epub 2004 Apr 23.

PY - 2004

Y1 - 2004

N2 - NR1/NR2D NMDA receptors display unusually slow deactivation kinetics which may be critical for their role as extrasynaptic receptors. A threonine to alanine point mutation has been inserted at amino acid position 692 of the NR2D subunit (T692A). Recombinant NR1a/NR2D(T692A) NMDA receptors have been expressed in Xenopus laevis oocytes and their pharmacological and single-channel properties examined using two-electrode voltage-clamp and patch-clamp recording techniques. Glutamate dose-response curves from NR1a/NR2D(T692A) receptor channels produced an approximately 1600-fold reduction in glutamate potency compared to wild-type NR1a/NR2D receptors. There was no change in Hill slopes or gross reduction in mean maximal currents recorded in oocytes expressing either wild-type or mutant receptors. The mutation did not affect the potency of the co-agonist glycine. The shifts in potency produced by NR2D(T692A) containing receptors when activated by other glutamate-site agonists such as aspartate or NMDA were 30- to 60-fold compared to wild-type. Single-channel conductance levels of NR1a/NR2D(T692A) mutant receptors were indistinguishable from wild-type NR2D-containing channels. Additionally NR1a/NR2D(T692A) receptors showed the transitional asymmetry that is characteristic of NR2D-containing NMDA receptors. Rapid applications of glutamate on outside-out patches containing NR1a/NR2D(T692A) receptors produced macroscopic current deactivations that were about 60-fold faster than wild-type NR1a/NR2D receptors. Our results suggest that this conserved threonine residue plays a crucial role in ligand binding to NMDA NR2 receptor subunits and supports the idea that the slow decay kinetics associated with NR1a/NR2D NMDA receptors can be explained by the slow dissociation of glutamate from this NMDA receptor subtype.

AB - NR1/NR2D NMDA receptors display unusually slow deactivation kinetics which may be critical for their role as extrasynaptic receptors. A threonine to alanine point mutation has been inserted at amino acid position 692 of the NR2D subunit (T692A). Recombinant NR1a/NR2D(T692A) NMDA receptors have been expressed in Xenopus laevis oocytes and their pharmacological and single-channel properties examined using two-electrode voltage-clamp and patch-clamp recording techniques. Glutamate dose-response curves from NR1a/NR2D(T692A) receptor channels produced an approximately 1600-fold reduction in glutamate potency compared to wild-type NR1a/NR2D receptors. There was no change in Hill slopes or gross reduction in mean maximal currents recorded in oocytes expressing either wild-type or mutant receptors. The mutation did not affect the potency of the co-agonist glycine. The shifts in potency produced by NR2D(T692A) containing receptors when activated by other glutamate-site agonists such as aspartate or NMDA were 30- to 60-fold compared to wild-type. Single-channel conductance levels of NR1a/NR2D(T692A) mutant receptors were indistinguishable from wild-type NR2D-containing channels. Additionally NR1a/NR2D(T692A) receptors showed the transitional asymmetry that is characteristic of NR2D-containing NMDA receptors. Rapid applications of glutamate on outside-out patches containing NR1a/NR2D(T692A) receptors produced macroscopic current deactivations that were about 60-fold faster than wild-type NR1a/NR2D receptors. Our results suggest that this conserved threonine residue plays a crucial role in ligand binding to NMDA NR2 receptor subunits and supports the idea that the slow decay kinetics associated with NR1a/NR2D NMDA receptors can be explained by the slow dissociation of glutamate from this NMDA receptor subtype.

KW - Animals Binding Sites/physiology Glutamic Acid/pharmacology Ion Channel Gating/drug effects/physiology Ligands Mutagenesis, Site-Directed Oocytes/physiology Patch-Clamp Techniques Protein Structure, Tertiary Rats Receptors, N-Methyl-D-Aspartate/agonists/g

M3 - Article

VL - 558

SP - 45

EP - 58

JO - Journal of Physiology

JF - Journal of Physiology

SN - 0022-3751

IS - Pt 1

ER -