Brain-derived neurotrophic factor increases Ca2+/calmodulin-dependent protein kinase 2 activity in hippocampus
Brain-derived neurotrophic factor increases Ca2+/calmodulin-dependent protein kinase 2 activity in hippocampus
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DOI：
10.1074/jbc.272.39.24133
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发表时间：
1997-09-26
影响因子：
4.8
通讯作者：
Lamour, Y
中科院分区：
文献类型：
--
作者：
Blanquet, PR;Lamour, Y
在这里，我们发现脑源性神经营养因子 (BDNF) 刺激大鼠海马切片中 Ca2+/钙调蛋白依赖性蛋白激酶 2 (CaMK2) 的磷酸化及其激酶活性。此外，我们发现：(i) BDNF 作用的时间过程并不伴随免疫印迹检测到的 CaMK2 α 和 β 亚基谱的变化； (ii)用碱性磷酸酶处理溶解的CaMK2和用蛋白磷酸酶1处理免疫沉淀的CaMK2均逆转激酶的磷酸化和激活； (iii)磷脂酶C抑制剂D609和1,2-双-(邻氨基苯氧基)乙烷-N,N,N'',N',-四乙酸四(乙酰氧基甲基)酯或8-(二乙氨基)辛基-3, 4,5-三甲氧基苯甲酸酯的细胞内Ca2+螯合，但不遗漏Ca2+或 EGTA 螯合 Ca2+，消除 BDNF 对 CaMK2 磷酸化和激活的刺激作用。这些结果强烈表明，BDNF 通过刺激磷脂酶 C 和随后的细胞内 Ca2+ 动员，增加 CaMK2 转化为其活性自磷酸化形式，但不增加其浓度。
Here we show that brain-derived neurotrophic factor (BDNF) stimulates both the phosphorylation of the Ca2+/calmodulin-dependent protein kinase 2 (CaMK2) and its kinase activity in rat hippocampal slices. In addition, we find that: (i) the time course of BDNF action is not accompanied by a change in the spectrum of either alpha- and beta-subunits of CaMK2 detected by immunoblotting; (ii) both treatment of solubilized CaMK2 with alkaline phosphatase and treatment of immunoprecipitated CaMK2 with protein phosphatase 1 reverse phosphorylation and activation of the kinase; (iii) phospholipase C inhibitor D609 and intracellular Ca2+ chelation by 1,2-bis-(o-aminophenoxy)ethane-N,N,N '',N',-tetracetic acid tetra(acetoxymethyl) ester or 8-(diethylamino) octyl-3, 4,5-trimethoxybenzoate but not omission of Ca2+ or Ca2+ chelation by EGTA, abolish the stimulatory effect of BDNF on phosphorylation and activation of CaMK2. These results strongly suggest that the conversion of CaMK2 into its active, autophosphorylated form, but not its concentration, is increased by BDNF via stimulation of phospholipase C and subsequent intracellular Ca2+ mobilization.