Interaction of the N-methyl-D-aspartic acid receptor NR2D subunit with the c-Abl tyrosine kinase

Robert T. Glover, Maria Angiolieri, Steven Kelly, Daniel T. Monaghan, Jean Y.J. Wang, Thomas E. Smithgall, Amy L. Buller

Research output: Contribution to journalArticle

18 Citations (Scopus)

Abstract

The COOH-terminal domain of the NR2D subunit of the NMDA receptor contains proline-rich regions that show striking homology to sequences known to bind to Src homology 3 (SH3) domains. To determine whether the proline- rich region of the NR2D subunit interacts with specific SH3 domains, in vitro SH3 domain binding assays were performed. A proline-rich fragment of the NR2D subunit (2D866-1064) bound to the Abl SH3 domain but not to the SH3 domains from Src, Fyn, Grb2, GAP, or phospholipase C-γ (PLCγ). Co- immunoprecipitation of NR2D with Abl suggests stable association of NR2D and Abl in transfected cells. The SH3 domain plays an important role in the negative regulation of Abl kinase activity. To determine whether the interaction of NR2D with the Abl SH3 domain alters Abl kinase activity, Abl was expressed alone or with NR2D in 293T cells. Autophosphorylation of Abl was readily observed when Abl was expressed alone. However, co-expression of Abl with 2D866-1064 or full-length NR2D inhibited autophosphorylation. 2D866-1064 did not inhibit ΔSH3 Abl, indicating a requirement for the Abl SH3 domain in the inhibitory effect. Similarly, 2D866-1064 did not inhibit the catalytic activity of Abl-PP, which contains two point mutations in the SH2-kinase linker domain that release the negative kinase regulation by the SH3 domain. In contrast, the full-length NR2D subunit partially inhibited the autokinase activity of both ΔSH3 Abl and Abl-PP, suggesting that NR2D and Abl may interact at multiple sites. Taken together, the data in this report provide the first evidence for a novel inhibitory interaction between the NR2D subunit of the NMDA receptor and the Abl tyrosine kinase.

Original languageEnglish (US)
Pages (from-to)12725-12729
Number of pages5
JournalJournal of Biological Chemistry
Volume275
Issue number17
DOIs
StatePublished - Apr 28 2000

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src Homology Domains
N-Methylaspartate
Protein-Tyrosine Kinases
Phosphotransferases
Proline
N-Methyl-D-Aspartate Receptors
Type C Phospholipases
Assays
Catalyst activity
Association reactions
aspartic acid receptor
HEK293 Cells
Sequence Homology
Immunoprecipitation
Point Mutation

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology
  • Cell Biology

Cite this

Glover, R. T., Angiolieri, M., Kelly, S., Monaghan, D. T., Wang, J. Y. J., Smithgall, T. E., & Buller, A. L. (2000). Interaction of the N-methyl-D-aspartic acid receptor NR2D subunit with the c-Abl tyrosine kinase. Journal of Biological Chemistry, 275(17), 12725-12729. https://doi.org/10.1074/jbc.275.17.12725

Interaction of the N-methyl-D-aspartic acid receptor NR2D subunit with the c-Abl tyrosine kinase. / Glover, Robert T.; Angiolieri, Maria; Kelly, Steven; Monaghan, Daniel T.; Wang, Jean Y.J.; Smithgall, Thomas E.; Buller, Amy L.

In: Journal of Biological Chemistry, Vol. 275, No. 17, 28.04.2000, p. 12725-12729.

Research output: Contribution to journalArticle

Glover, RT, Angiolieri, M, Kelly, S, Monaghan, DT, Wang, JYJ, Smithgall, TE & Buller, AL 2000, 'Interaction of the N-methyl-D-aspartic acid receptor NR2D subunit with the c-Abl tyrosine kinase', Journal of Biological Chemistry, vol. 275, no. 17, pp. 12725-12729. https://doi.org/10.1074/jbc.275.17.12725
Glover, Robert T. ; Angiolieri, Maria ; Kelly, Steven ; Monaghan, Daniel T. ; Wang, Jean Y.J. ; Smithgall, Thomas E. ; Buller, Amy L. / Interaction of the N-methyl-D-aspartic acid receptor NR2D subunit with the c-Abl tyrosine kinase. In: Journal of Biological Chemistry. 2000 ; Vol. 275, No. 17. pp. 12725-12729.
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abstract = "The COOH-terminal domain of the NR2D subunit of the NMDA receptor contains proline-rich regions that show striking homology to sequences known to bind to Src homology 3 (SH3) domains. To determine whether the proline- rich region of the NR2D subunit interacts with specific SH3 domains, in vitro SH3 domain binding assays were performed. A proline-rich fragment of the NR2D subunit (2D866-1064) bound to the Abl SH3 domain but not to the SH3 domains from Src, Fyn, Grb2, GAP, or phospholipase C-γ (PLCγ). Co- immunoprecipitation of NR2D with Abl suggests stable association of NR2D and Abl in transfected cells. The SH3 domain plays an important role in the negative regulation of Abl kinase activity. To determine whether the interaction of NR2D with the Abl SH3 domain alters Abl kinase activity, Abl was expressed alone or with NR2D in 293T cells. Autophosphorylation of Abl was readily observed when Abl was expressed alone. However, co-expression of Abl with 2D866-1064 or full-length NR2D inhibited autophosphorylation. 2D866-1064 did not inhibit ΔSH3 Abl, indicating a requirement for the Abl SH3 domain in the inhibitory effect. Similarly, 2D866-1064 did not inhibit the catalytic activity of Abl-PP, which contains two point mutations in the SH2-kinase linker domain that release the negative kinase regulation by the SH3 domain. In contrast, the full-length NR2D subunit partially inhibited the autokinase activity of both ΔSH3 Abl and Abl-PP, suggesting that NR2D and Abl may interact at multiple sites. Taken together, the data in this report provide the first evidence for a novel inhibitory interaction between the NR2D subunit of the NMDA receptor and the Abl tyrosine kinase.",
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