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Roles of Tyrosine Phosphorylation in The Development and Plasticity of The Central Nervous System

酪氨酸磷酸化在中枢神经系统发育和可塑性中的作用

基本信息

批准号：
11308031
负责人：
YAMAMOTO Tadashi
金额：
$ 24.9万
依托单位：
The University of Tokyo
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (A)
财政年份：
1999
资助国家：
日本
起止时间：
1999 至 2001
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-11308031/
关键词：
Novel tyrosine kinases Central nervous system RT-PCR in silico screening Signal transduction ALK Lyn Filter kinase assay チロシンキナーゼチロシンフォスファターゼシナプス伝達神経回路網 NMDA受容体 Srcファミリー SSH法 ERBB2 LYN フィルターキナーゼアッセイ

项目摘要

We performed three types of studies that are (1) the search of the novel tyrosine kinases expressed in the central nervous system, (2) identification of proteins that are respectively involved in Lyn and ALK tyrosine kinases-mediated signaling pathways in the central nervous system, (3) characterization of a protein-tyrosine phosphatase PTPMEG that associates with ionotropic glutamate receptors. The followings are the summary of the study.(1) By performing differential screening of RT-PCR products and in silico screening, we identified novel kinases that included the one termed BREK (brain enriched kinase). We also identified a homologue of BREK termed BREK2. BREK and BREK2 are similar to AATYK that has been reported to be a tyrosine kinase involved in regulation of apoptotic death of neurons. Unlike AATYK, both BREK and BREK2 are dual specificity kinases showing strong serine/threonine and weak tyrosine kinase activities. Expression of BREK is high in the central nervous system, such as in cerebral cortex and olfactory bulb. BREK becomes phosphorylated upon NGF stimulation of cultured neuronal cells, suggesting that BREK is involved in neurite extension.(2) By employing filter kinase assay and yeast two-hybrid screening, we identified BANK and SNT2 as substrates of Lyn and ALK, respectively. BANK is an adaptor protein that links Lyn to IP_3 receptor (IP_3R). We showed that formation of Lyn/BANK/IP_3R resulted in tyrosine phosphorylation of IP_3R, which is known to upregulate the channel activity of IP_3R. Regarding SNT2, we showed that SNT2 was critically important for ALK-mediated signaling.(3) We also identified various signaling molecules that were associated with ionotropic glutamate receptors. One of them is PTPMEG that associates with GluRd2 and NMDA receptor subunits NR2A. PTPMEG is expressed high in the thalamus and affects the level of tyrosine phosphorylation of NR2A, suggesting that PTPMEG is involved in controlling synaptic plasticity.

我们进行了三种类型的研究，即（1）寻找在中枢神经系统中表达的新酪氨酸激酶，（2）鉴定分别参与中枢神经系统中林恩和ALK酪氨酸激酶介导的信号通路的蛋白，（3）表征与离子型谷氨酸受体相关的蛋白酪氨酸磷酸酶PTPMEG。以下是研究的总结。(1)通过对RT-PCR产物进行差异筛选和计算机筛选，我们鉴定了新的激酶，包括称为BREK（脑富集激酶）的激酶。我们还鉴定了BREK的同源物，称为BREK 2。BREK和BREK 2与AATYK相似，据报道AATYK是一种参与调节神经元凋亡性死亡的酪氨酸激酶。与AATYK不同，BREK和BREK 2都是显示强丝氨酸/苏氨酸和弱酪氨酸激酶活性的双特异性激酶。BREK在中枢神经系统如大脑皮层和嗅球中表达高。BREK在NGF刺激培养的神经元细胞后磷酸化，表明BREK参与神经突延伸。(2)通过滤纸激酶试验和酵母双杂交筛选，我们确定BANK和SNT 2分别为林恩和ALK的底物。BANK是连接林恩和IP_3受体（IP_3R）的接头蛋白。我们发现，林恩/BANK/IP_3 R的形成导致IP_3 R的酪氨酸磷酸化，这是已知的上调IP_3 R的通道活性。关于SNT 2，我们发现SNT 2对ALK介导的信号传导至关重要。(3)我们还鉴定了与离子型谷氨酸受体相关的各种信号分子。其中之一是PTPMEG，其与GluRd 2和NMDA受体亚基NR 2A相关。PTPMEG在丘脑中高表达，并影响NR 2A的酪氨酸磷酸化水平，表明PTPMEG参与控制突触可塑性。