Post-mortem interval effects on the phosphorylation of signaling proteins
Post-mortem interval effects on the phosphorylation of signaling proteins
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DOI：
10.1038/sj.npp.1300112
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发表时间：
2003-06-01
影响因子：
7.6
通讯作者：
Chen, G
中科院分区：
文献类型：
--
作者：
Li, JL;Gould, TD;Chen, G
死后的脑组织提供了一个独特的机会，揭示基因或蛋白质参与的病理生理学的神经精神疾病。蛋白质磷酸化是细胞内信号通路中常见的蛋白质修饰，其影响蛋白质的分布和功能，并且已被假设在主要神经精神疾病的病理生理学和治疗中具有重要意义。因此，我们有兴趣确定参与细胞信号传导的蛋白质磷酸化形式的稳定性。针对磷酸酪氨酸、磷酸苏氨酸和磷酸PKA底物的抗体用于检查PMI对磷酸化形式的蛋白质的一般量的影响。ERK、JNK、RSK、CREB和ATF-2的磷酸化特异性抗体用于测试PMI对特定蛋白质的作用，已知这些蛋白质的功能显著受磷酸化调节。我们发现，PMI迅速降低了磷酸化状态的蛋白质水平，也降低了某些蛋白质的总水平。在4 ℃和室温下储存的样品中，在额叶皮层和海马中都观察到了PMI效应。因此，似乎依赖于蛋白质磷酸化状态的测量（如二维凝胶电泳和功能测定）对PMI非常敏感。
Post-mortem brain tissue provides a unique opportunity to uncover the genes or proteins involved in the pathophysiology of neuropsychiatric disorders. Protein phosphorylation is a common protein modification within intracellular signaling pathways that affects the distribution and function of protein, and has been hypothesized to be of major importance in both the pathophysiology and treatment of major neuropsychiatric disorders. Thus, we were interested in ascertaining the stability of the phosphorylated forms of proteins that are involved in cellular signaling. Antibodies against phospho-tyrosine, phospho-threonine, and phospho-PKA substrates were used to examine the PMI effects on the general amounts of proteins in their phosphorylated form. Phospho-specific antibodies for ERK, JNK, RSK, CREB, and ATF-2 were used to test the effects of PMI on specific proteins whose functioning are known to be regulated markedly by phosphorylation. We found that PMI rapidly decreased the levels of proteins in their phosphorylated states and also decreased the total levels of certain proteins. The PMI effects were observed in the samples stored at both 4degreesC and room temperature, in both frontal cortex and hippocampus. Thus, it appears that measurements (such as two-dimensional gel electrophoresis and functional assays) that rely on the phosphorylation state of proteins would be extremely sensitive to PMI.