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机构地区:[1]北京师范大学生命科学学院基因工程药物及生物技术北京市重点实验室,北京100875 [2]北京师范大学生命科学学院细胞增殖及调控生物学教育部重点实验室,北京100875 [3]北京师范大学生命科学学院抗性基因与分子发育北京市重点实验室,北京100875
出 处:《科技导报》2012年第31期73-79,共7页Science & Technology Review
基 金:国家自然科学基金项目(30971470);北京市自然科学基金项目(5122017);细胞增殖及调控生物学教育部重点实验室开放基金项目(201101);教育部2011年度大学生创新创业训练计划项目;抗性基因与分子发育北京市重点实验室开放基金项目(01102)
摘 要:蛋白质磷酸化是由蛋白质激酶催化的磷酸基转移反应,是最常见、最重要的蛋白质翻译后修饰方式之一,是一种普遍的生命活动调节方式,在细胞信号转导过程中起重要作用。本文介绍了蛋白质磷酸化修饰的主要类型与功能、磷酸化蛋白的鉴定及磷酸化位点的预测等方面研究进展,并着重介绍了一些灵敏度高、特异性强的以同位素标记、免疫印迹-化学发光法等作为核心的磷酸化蛋白质分析方案。Western blot方法被证明是鉴别磷蛋白的灵敏、特异方法,而Nano Pro100/1000超微量蛋白分析系统等又在此基础上加以改善。蛋白磷酸化分析工具和软件的发展也很迅猛。Phosphorylation is one of the most important post-translational protein modifications, which is related to the regulation of many activities of life. As a sort of reaction catalyzed by protein kinases, it functions significantly in cell signaling. To keep up with the situation that protein phosphorylation analysis and phosphorylated site identification have been a focus of the current proteomic study, and benefit the functional dissection on various protein phosphorylation reactions, this article introduces the progress in the studies of main types and functions of protein phosphorylation, the identification of the phosphopeptides, the determination and prediction of specific phosphorylation sites. As a focal point, protocols based on isotope labeling, chemilumineseence, etc, are elucidated. Western blotting has been proven sensitive and specific in the identification of phosphoproteins, based on which, there have appeared some improved approaches, such as NanoProl00/1000 ultramicro-protein analysis system. Phosphorylation analytic tools and softwares are being swiftly developing. According to numerous researches, protein phosphorylation/dephosphorylation modifications are important with many aspects of meaning. Controlling the cellular endogenous enzymic "activities', protein phosphorylation/dephosphorylation are more able to rapidly react to external stimulus than enzymic re-synthesis/decomposition. A seriesof phosphorylation/dephosphorylation reactions in different tissues and with different temporal and spatial characteristics ensure the sustained responses of cells to external signals, and play a role in the cascade amplifying for the external signals. Protein phosphorylation/dephosphorylation study will continue to be a hotspot in the exporation of life activity regulatory mechanism.
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