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
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
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DOI：
10.1074/jbc.274.9.5782
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发表时间：
1999-02-26
影响因子：
4.8
通讯作者：
Saya, H
中科院分区：
文献类型：
--
作者：
Masuko, N;Makino, K;Saya, H
NE-dlg/SAP 102是一种神经元和内分泌组织特异性膜相关鸟苷酸激酶家族蛋白，已知其通过其PDZ（PSD-95/Dlg/ZO-1）结构域结合N-甲基-D-天冬氨酸受体2B（NR 2B）的C末端。NE-dlg/SAP 102和NR 2B在培养的大鼠海马神经元中共定位于突触部位，并且它们的表达与突触发生的开始平行地增加，我已经确定NE-dlg/SAP 102以Ca 2+依赖的方式与钙调素相互作用。钙调素的结合位点已被确定为位于NE-dlg/SAP 102的src同源性3（SH 3）结构域周围的推定的碱性cr-螺旋区。利用表面等离子体共振测量系统，我们检测到重组NE-dlg/SAP 102与固定化钙调素的特异性结合，Kd值为44 nM。然而，Ca ~（2+）/钙调素与NE-dlg/SAP 102的结合并不调节NE-dlg/SAP 102的PDZ结构域与大鼠NR 2B的C-末端之间的相互作用。我们还通过双杂交筛选确定了NE-dlg/SAP 102的钙调素结合位点附近的区域与PSD-95/SAP 90的GUK样结构域相互作用。Pull down实验表明，NE-dlg/SAP 102在Ca ~（2+）和钙调素存在下能与PSD-95/SAP 90相互作用。这些发现表明，Ca 2 +/钙调素调节神经元膜相关的鸟苷酸激酶蛋白的相互作用，并调节在中央突触的神经递质受体的集群。
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, me have identified that NE-dlg/SAP102 interacts with calmodulin in a Ca2+-dependent manner. The binding site for calmodulin has been determined to he at the putative basic cr-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 interacts 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.