Regulation of phosphorylation at the postsynaptic density during different activity states of Ca2+/calmodulin-dependent protein kinase II.
Regulation of phosphorylation at the postsynaptic density during different activity states of Ca2+/calmodulin-dependent protein kinase II.
复制标题
DOI：
10.1016/j.bbrc.2009.10.167
复制
发表时间：
2010-01-01
影响因子：
3.1
通讯作者：
Jaffe, Howard
中科院分区：
文献类型：
--
作者：
Dosemeci, Ayse;Jaffe, Howard
钙/钙调蛋白依赖性蛋白激酶II(CaMKII)是突触后密度(PSD)中含量最丰富的一种蛋白，被认为参与了对突触特性的活性诱导调节。CaMKII在有钙离子存在的情况下与钙调蛋白结合时被激活，一旦在T-286/7上自动磷酸化，在没有钙离子(自主形式)的情况下仍保持活性。在本研究中，我们使用定量质谱学策略(ITRAQ)来识别在CaMKII激活时被磷酸化的PSD组分上的位点。在CaMKII处于(1)大部分不活跃(基础状态)，(2)在有钙离子存在时活跃，(3)在无钙离子存在时活跃的情况下，我们监测了分离的PSD的磷酸化。通过确认前面描述的CaMKII的自磷酸化特性，验证了定量策略。在钙离子存在和不存在的情况下，激活的CaMKII对PSD主要成分的磷酸化效果各不相同。最值得注意的是，在没有钙离子的情况下，自主活性对SynGAP上三个残基的磷酸化更有效。几个PSD支架蛋白在激活CaMKII时被磷酸化。采用的策略首次在SAPAPs和Shanks上鉴定了CaMKII调控的位点，包括SAPAP1、SAPAP2和SAPAP3的C-末端附近的三个保守的丝氨酸残基。CaMKII参与PSD支架蛋白的磷酸化，提示在活性诱导的PSD结构重组中发挥作用。
Ca2+/calmodulin dependent protein kinase II (CaMKII), the most abundant kinase at the postsynaptic density (PSD), is expected to be involved in activity-induced regulation of synaptic properties. CaMKII is activated when it binds calmodulin in the presence of Ca2+ and, once autophosphorylated on T-286/7, remains active in the absence of Ca2+ (autonomous form). In the present study we used a quantitative mass spectrometric strategy (iTRAQ) to identify sites on PSD components phosphorylated upon CaMKII activation. Phosphorylation in isolated PSDs was monitored under conditions where CaMKII is: (1) mostly inactive (basal state), (2) active in the presence of Ca2+ and (3) active in the absence of Ca2+. The quantification strategy was validated through confirmation of previously described autophosphorylation characteristics of CaMKII. The effectiveness of phosphorylation of major PSD components by the activated CaMKII in the presence and absence of Ca2+ varied. Most notably, autonomous activity in the absence of Ca2+ was more effective in the phosphorylation of three residues on SynGAP. Several PSD scaffold proteins were phosphorylated upon activation of CaMKII. The strategy adopted allowed the identification, for the first time, of CaMKII-regulated sites on SAPAPs and Shanks, including three conserved serine residues near the C-termini of SAPAP1, SAPAP2 and SAPAP3. Involvement of CaMKII in the phosphorylation of PSD scaffold proteins suggests a role in activity-induced structural re-organization of the PSD.
登录
查看更多内容
影响因子：
7
作者：
Trinidad, Jonathan C.;Thalhammer, Agnes;Burlingame, Alma L.
通讯作者：
Burlingame, Alma L.
影响因子：
4.7
作者：
Thalhammer A;Trinidad JC;Burlingame AL;Schoepfer R
通讯作者：
Schoepfer R
影响因子：
4.8
作者：
Collins, MO;Yu, L;Grant, SGN
通讯作者：
Grant, SGN
影响因子：
7
作者：
Trinidad, JC;Specht, CG;Burlingame, AL
通讯作者：
Burlingame, AL
影响因子：
16.2
作者：
Naisbitt, S;Kim, E;Sheng, M
通讯作者：
Sheng, M