Interaction of NE-dlg/SAP102, a neuronal and endocrine tissue-specific membrane-associated guanylate kinase protein, with calmodulin and PSD- 95/SAP90: A possible regulatory role in molecular clustering at synaptic sites

Norio Masuko, Keishi Makino, Hiroaki Kuwahara, Kohji Fukunaga, Tamotsu Sudo, Norie Araki, Hideyuki Yamamoto, Yuji Yamada, Eishichi Miyamoto, Hideyuki Saya

Research output: Contribution to journalArticle

76 Citations (Scopus)

Abstract

NE-dlg/SAP102, a neuronal and endocrine tissue-specific membrane- associated guanylate kinase family protein, is known to bind to C-terminal ends of N-methyl-D-aspartate receptor 2B (NR2B) through its PDZ (PSD- 95/Dlg/ZO-1) domains. NE-dlg/SAP102 and NR2B colocalize at synaptic sites in cultured rat hippocampal neurons, and their expressions increase in parallel with the onset of synaptogenesis. We have identified that NE-dlg/SAP102 interacts with calmodulin in a Ca2+-dependent manner. The binding site for calmodulin has been determined to lie at the putative basic α-helix region located around the src homology 3 (SH3) domain of NE-dlg/SAP102. Using a surface plasmon resonance measurement system, we detected specific binding of recombinant NE-dlg/SAP102 to the immobilized calmodulin with a K(d) value of 44 nM. However, the binding of Ca2+/calmodulin to NE-dlg/SAP102 did not modulate the interaction between PDZ domains of NE-dlg/SAP102 and the C- terminal end of rat NR2B. We have also identified that the region near the calmodulin binding site of NE-dlg/SAP102 interact with the GUK-like domain of PSD-95/SAP90 by two-hybrid screening. Pull down assay revealed that NE- dlg/SAP102 can interact with PSD-95/SAP90 in the presence of both Ca2+ and calmodulin. These findings suggest that the Ca2+/calmodulin modulates interaction of neuronal membrane-associated guanylate kinase proteins and regulates clustering of neurotransmitter receptors at central synapses.

Original languageEnglish
Pages (from-to)5782-5790
Number of pages9
JournalJournal of Biological Chemistry
Volume274
Issue number9
DOIs
Publication statusPublished - 1999 Feb 26
Externally publishedYes

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology
  • Cell Biology

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