The N‐Methyl‐d‐Aspartate Receptor Subunits NR2A and NR2B Bind to the SH2 Domains of Phospholipase C‐γ
The N‐Methyl‐d‐Aspartate Receptor Subunits NR2A and NR2B Bind to the SH2 Domains of Phospholipase C‐γ
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N-甲基-d-天冬氨酸受体亚基 NR2A 和 NR2B 与磷脂酶 C-γ 的 SH2 结构域结合
DOI：
10.1046/j.1471-4159.1997.69020623.x
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发表时间：
1997
影响因子：
4.7
通讯作者：
N. Bissoon
中科院分区：
文献类型：
--
作者：
J. Gurd;N. Bissoon
摘要：最近发现NMDA受体在酪氨酸上被磷酸化。为了探讨突触后细胞中N-甲基-D-天冬氨酸受体酪氨酸磷酸化与信号通路之间的可能联系，我们研究了N-甲基-D-天冬氨酸受体亚单位与磷脂酶C-γ的SH2区结合(PLC-γ)与酪氨酸磷酸化之间的关系。含有PLC-γ的N-和C-近端SH2结构域的谷胱甘肽S转移酶融合蛋白与谷胱甘肽-琼脂糖凝胶结合，并与突触连接蛋白和糖蛋白反应。酪氨酸磷酸化的PSD-GP180被鉴定为NMDA受体的NR2B亚单位，以磷酸化依赖的方式结合到SH2-琼脂糖珠上。用针对单个NMDA受体亚基的抗体进行免疫印迹分析表明，NR2A和NR2B亚基都与SH2-琼脂糖结合。没有与缺乏相关SH2结构域的GST-琼脂糖结合，表明结合是SH2结构域所特有的。突触连接与三磷酸腺苷孵育后受体亚基结合增加，而外源性蛋白酪氨酸磷酸酶处理突触连接后受体亚基结合减少。免疫沉淀实验证实，NR2A和NR2B在酪氨酸上发生了磷酸化，而且与ATP孵育后，每个亚基的酪氨酸磷酸化程度都增加了。结果表明，NMDA受体亚基NR2a和NR2B将结合到PLC-γ的SH2结构域，分离的突触连接含有内源性蛋白酪氨酸激酶(S)，可以同时磷酸化NR2a和NR2B受体亚基，提示酪氨酸磷酸化的NMDa受体与含有SH2结构域的蛋白质相互作用可能有助于将其与突触后细胞中的信号通路联系起来。
Abstract: The NMDA receptor has recently been found to be phosphorylated on tyrosine. To assess the possible connection between tyrosine phosphorylation of the NMDA receptor and signaling pathways in the postsynaptic cell, we have investigated the relationship between tyrosine phosphorylation and the binding of NMDA receptor subunits to the SH2 domains of phospholipase C‐γ (PLC‐γ). A glutathione S‐transferase (GST) fusion protein containing both the N‐ and the C‐proximal SH2 domains of PLC‐γ was bound to glutathione‐agarose and reacted with synaptic junctional proteins and glycoproteins. Tyrosine‐phosphorylated PSD‐GP180, which has been identified as the NR2B subunit of the NMDA receptor, bound to the SH2‐agarose beads in a phosphorylation‐dependent fashion. Immunoblot analysis with antibodies specific for individual NMDA receptor subunits showed that both NR2A and NR2B subunits bound to the SH2‐agarose. No binding occurred to GST‐agarose lacking an associated SH2 domain, indicating that binding was specific for the SH2 domains. The binding of receptor subunits increased after the incubation of synaptic junctions with ATP and decreased after treatment of synaptic junctions with exogenous protein tyrosine phosphatase. Immunoprecipitation experiments confirmed that NR2A and NR2B were phosphorylated on tyrosine and further that tyrosine phosphorylation of each of the subunits was increased after incubation with ATP. The results demonstrate that NMDA receptor subunits NR2A and NR2B will bind to the SH2 domains of PLC‐γ and that isolated synaptic junctions contain endogenous protein tyrosine kinase(s) that can phosphorylate both NR2A and NR2B receptor subunits, and suggest that interaction of the tyrosine‐phosphorylated NMDA receptor with proteins that contain SH2 domains may serve to link it to signaling pathways in the postsynaptic cell.
影响因子：
56.9
作者：
GRANT, SGN;ODELL, TJ;KANDEL, ER
通讯作者：
KANDEL, ER
影响因子：
56.9
作者：
BADING, H;GREENBERG, ME
通讯作者：
GREENBERG, ME
神经生长因子通过与 trk 原癌基因产物相关的激酶活性快速刺激磷脂酶 C-gamma 1 的酪氨酸磷酸化。
DOI：
10.1073/pnas.88.13.5650
发表时间：
1991
影响因子：
11.1
作者：
Vetter,ML;Martin-Zanca,D;Parada,LF;Bishop,JM;Kaplan,DR
通讯作者：
Kaplan,DR