| cdk-5过度表达对神经细丝磷酸化的影响 |
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| 引用本文： | 陈娟,冯友梅,周洁,郑伟,李宏莲,许亮,王建枝.cdk-5过度表达对神经细丝磷酸化的影响[J].医学分子生物学杂志,2004,1(2):69-72. |
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| 作者姓名： | 陈娟 冯友梅 周洁 郑伟 李宏莲 许亮 王建枝 |
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| 作者单位： | 华中科技大学同济医学院基础医学院生物化学与分子生物学系,武汉市,430030;华中科技大学同济医学院基础医学院病理生理学系,武汉市,430030;华中科技大学生命科学院生物物理与生物化学研究所,武汉市,430030 |
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| 基金项目： | 国家自然科学基金(30100057,30170221) |
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| 摘 要： | 目的在细胞水平研究cdk-5过度表达对神经细丝磷酸化的影响。方法 用脂质体转染技术将cdk-5基因转入N2a细胞株中,并建立稳定表达cdk-5的N2a/cdk-5细胞株,免疫沉淀法及酶活性测定检测cdk-5活性,免疫荧光和免疫印迹技术检测cdk-5的表达和神经细丝的磷酸化状态。结果 在N2a细胞株转染组中,cdk-5表达增加,并使抗体SMI31显色增强,SMI32显色减弱,提示神经细丝被过度磷酸化。与此同时,cdk-5酶活性较未转染组提高3.5倍。结论提示细胞水平的cdk-5过度表达会导致cdk-5过度激活和神经细丝过度磷酸化,而过度磷酸化的神经细丝可能参与了阿尔茨海默病的病理过程。
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| 关 键 词： | 细胞周期依赖性蛋白激酶5(cdk-5) 神经细丝 阿尔茨海默病 |
| 修稿时间： | 2004年2月25日 |
The Effect of cdk-5 Overexpression on Neurofilament Phosphorylation |
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| CHEN Juan,FENG Youmei,ZHOU Jie,ZHENG Wei,LI Honglian,XU Liang,WANG Jianzhi.The Effect of cdk-5 Overexpression on Neurofilament Phosphorylation[J].Journal of Medical Molecular Biology,2004,1(2):69-72. |
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| Authors: | CHEN Juan FENG Youmei ZHOU Jie ZHENG Wei LI Honglian XU Liang WANG Jianzhi |
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| Institution: | CHEN Juan,FENG Youmei,ZHOU Jie,ZHENG Wei,LI Honglian,XU Liang,WANG Jianzhi Department of Biochemistry and Molecular Biology,Tongji Medical College,Huazhong University of Science and Technology,Wuhan,430030,China
Department of Pathophysiology,Tongji Medical College,Huazhong University of Science and Technology,Wuhan,430030,China
Institute of Biophysics and Biochemistry,Life Science and Technology College,Huazhong University of Science and Technology,Wuhan,430030,China |
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| Abstract: | Objective In this study, we investigated the effect of cyclin-dependent kinase (cdk-5) overexpression on neurofilament phosphorylation in cultured cells. Methods The cdk-5 was transfected into N2a cell line with cdk-5 by using Lipofectamine2000, the cdk-5 activity and neurofilament phosphorylation was detected by using immmunoprecipetation, cdk-5 activity assay, immun-ofluoresence and Western blots. Results It was found that overexpression of cdk-5 led to elevation of cdk-5 activity, which was 3. 5 times that of the control level. Overactivation of cdk-5 resulted in hyperphosphorylation of neurofilament. It was demonstrated that overexpression of cdk-5 caused reduced staining with antibody SMI32 and enhanced staining with antibody SMI31. Conclusion These results indicate that in vitro overexpression of cdk-5 could result in cdk-5 overactivation and neurofilament hyperphosphorylation, which might be the underlying mechanism of Alzheimer dis- |
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| Keywords: | cyclin-dependent kinase5 (cdk-5) neurofilament Alzheimer' s disease |
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