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Biology of 14-3 3 proteins regulating neuronal cell death

调节神经细胞死亡的 14-3 3 蛋白的生物学

基本信息

批准号：
18591143
负责人：
FUJII Katsunori
金额：
$ 2.57万
依托单位：
Chiba University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
2006
资助国家：
日本
起止时间：
2006 至 2007
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-18591143/
关键词：
14-3-3 proteins apoptosis Neuronal cell death molecular regulations Pediatric Neurology

项目摘要

We aimed in this study to elucidate the biological mechanisms regulating neuronal cell death and molecular interactions affecting the neurological diseases in children on both aspects, biological and clinical medicines.Detection of 14-3-3 protein in the CSF is a powerful tool for elucidating the underlying mechanisms of neurological disorders. There have been useful studies on 14-3-3 CSF protein detection in Creutzfeldt-Jakob disease and other neurological disorders, but none on cerebellar diseases. To evaluate detection of 14-3-3 protein in the CSF of patients with cerebellar diseases, we examined 14-3-3 protein in the CSF by immunoblotting in seven patients with cerebellar diseases, i.e., acute cerebellitis (2), acute cerebellar ataxia (2), and cerebellar atrophy (3). 14-3-3 protein isoforms were also identified by means of isoform-specific antibodies. 14-3-3 protein was detected in the CSF of six patients, the exception being one with acute cerebellar ataxia. Interestingly, only the … More 14-3-3 c isoform was detected throughout in 14-3-3 positive patients. Moreover, longitudinal analysis of 14-3-3 CSF protein in one patient with cerebellar atrophy showed that the 14-3-3 band density proportionally decreased when the cerebellar atrophy gradually progressed. We concluded that 14-3-3 protein in the CSF is a significant cerebellar destructive marker as well as one in other brain diseases, and the unique detection of 14-3-3 ε may indicate cerebellar involvement in the brain.Since 14-3-3 proteins are an intimate partner of apoptosis signal-regulating kinase 1 (ASK1), we examined in vitro protein kinse assay using recombinant ASK1 and 14-3-3 proteins. We clearly showed that ASK1 can phosphorylate 14-3-3 proteins with several portions of serine and threonine residues. Interestingly, phosphorylated 14-3-3 proteins diminished ASK1 autophosphorylation, suggesting that ASK1-14-3-3 proteins interaction may influence the kinetic activity of ASK1. As 14-3-3 proteins bind to ASK1 using a binding consensus motif, protein-protein interaction is supposed to be critical biological function on both ASK1 and 14-3-3 proteins. (305 words) Less

本研究旨在从生物学和临床医学两个方面阐明神经元死亡的生物学机制和影响儿童神经系统疾病的分子相互作用，脑脊液中14-3-3蛋白的检测是阐明神经系统疾病发病机制的有力工具。脑脊液中14-3-3蛋白的检测在克雅氏病和其他神经系统疾病中有一定的应用价值，但在小脑疾病中未见报道。为了评价小脑疾病患者CSF中14-3-3蛋白的检测，我们通过免疫印迹法检测了7名小脑疾病患者CSF中的14 -3-3蛋白，即，急性小脑炎（2）、急性小脑共济失调（2）和小脑萎缩（3）。14-3-3蛋白同种型也通过同种型特异性抗体鉴定。14-3-3蛋白在6例患者的CSF中检测到，除了1例急性小脑性共济失调患者。有趣的是，只有关于我们在14 - 3 -3阳性的患者中，14 -3-3 C异构体始终被检测到。此外，对1例小脑萎缩患者脑脊液14-3-3蛋白的纵向分析显示，随着小脑萎缩的逐渐进展，14-3-3带密度成比例地降低。我们的结论是，脑脊液中14-3-3蛋白是一个重要的小脑破坏性标志物，也是其他脑疾病的标志物，14-3-3 ε的独特检测可能表明小脑在脑中的参与，由于14-3-3蛋白是凋亡信号调节激酶1（ASK 1）的亲密伴侣，我们使用重组ASK 1和14-3-3蛋白检测了体外蛋白激酶测定。我们清楚地表明，ASK 1可以磷酸化14-3-3蛋白与几个部分的丝氨酸和苏氨酸残基。有趣的是，磷酸化的14-3-3蛋白减少了ASK 1的自磷酸化，这表明ASK 1 -14-3-3蛋白的相互作用可能影响ASK 1的动力学活性。由于14-3-3蛋白使用结合共有基序与ASK 1结合，因此蛋白质-蛋白质相互作用被认为是ASK 1和14-3-3蛋白的关键生物学功能。(305减