Characterization of Fyn-mediated tyrosine phosphorylation sites on GluRε2 (NR2B) subunit of the N-methyl-D-aspartate receptor

Takanobu Nakazawa, Shoji Komai, Tohru Tezuka, Chihiro Hisatsune, Hisashi Umemori, Kentaro Senba, Masayoshi Mishina, Toshiya Manabe, Tadashi Yamamoto

Research output: Contribution to journalArticle

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Abstract

The N-methyl-D-aspartate (NMDA) receptors play critical roles in synaptic plasticity, neuronal development, and excitotoxicity. Tyrosine phosphorylation of NMDA receptors by Src-family tyrosine kinases such as Fyn is implicated in synaptic plasticity. To precisely address the roles of NMDA receptor tyrosine phosphorylation, we identified Fyn-mediated phosphorylation sites on the GluRε2 (NR2B) subunit of NMDA receptors. Seven out of 25 tyrosine residues in the C-terminal cytoplasmic region of GluRε2 were phosphorylated by Fyn in vitro. Of these 7 residues, Tyr-1252, Tyr-1336, and Tyr-1472 in GluRε2 were phosphorylated in human embryonic kidney fibroblasts when co-expressed with active Fyn, and Tyr-1472 was the major phosphorylation site in this system. We then generated rabbit polyclonal antibodies specific to Tyr-1472-phosphorylated GluRε2 and showed that Tyr-1472 of GluRε2 was indeed phosphorylated in murine brain using the antibodies. Importantly, Tyr-1472 phosphorylation was greatly reduced in fyn mutant mice. Moreover, Tyr-1472 phosphorylation became evident when hippocampal long term potentiation started to be observed, and its magnitude became larger in murine brain. Finally, Tyr-1472 phosphorylation was significantly enhanced after induction of long term potentiation in the hippocampal CA1 region. These data suggest that Tyr-1472 phosphorylation of GluRε2 is important for synaptic plasticity.

Original languageEnglish
Pages (from-to)693-699
Number of pages7
JournalJournal of Biological Chemistry
Volume276
Issue number1
DOIs
Publication statusPublished - 2001 Jan 5
Externally publishedYes

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Phosphorylation
N-Methyl-D-Aspartate Receptors
Tyrosine
Neuronal Plasticity
Plasticity
Long-Term Potentiation
Brain
Hippocampal CA1 Region
src-Family Kinases
NR2B NMDA receptor
Antibodies
Fibroblasts
Rabbits
Kidney

ASJC Scopus subject areas

  • Biochemistry

Cite this

Characterization of Fyn-mediated tyrosine phosphorylation sites on GluRε2 (NR2B) subunit of the N-methyl-D-aspartate receptor. / Nakazawa, Takanobu; Komai, Shoji; Tezuka, Tohru; Hisatsune, Chihiro; Umemori, Hisashi; Senba, Kentaro; Mishina, Masayoshi; Manabe, Toshiya; Yamamoto, Tadashi.

In: Journal of Biological Chemistry, Vol. 276, No. 1, 05.01.2001, p. 693-699.

Research output: Contribution to journalArticle

Nakazawa, T, Komai, S, Tezuka, T, Hisatsune, C, Umemori, H, Senba, K, Mishina, M, Manabe, T & Yamamoto, T 2001, 'Characterization of Fyn-mediated tyrosine phosphorylation sites on GluRε2 (NR2B) subunit of the N-methyl-D-aspartate receptor', Journal of Biological Chemistry, vol. 276, no. 1, pp. 693-699. https://doi.org/10.1074/jbc.M008085200
Nakazawa, Takanobu ; Komai, Shoji ; Tezuka, Tohru ; Hisatsune, Chihiro ; Umemori, Hisashi ; Senba, Kentaro ; Mishina, Masayoshi ; Manabe, Toshiya ; Yamamoto, Tadashi. / Characterization of Fyn-mediated tyrosine phosphorylation sites on GluRε2 (NR2B) subunit of the N-methyl-D-aspartate receptor. In: Journal of Biological Chemistry. 2001 ; Vol. 276, No. 1. pp. 693-699.
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