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基于质谱技术的蛋白质组定量方法

期刊名称：生命的化学
时间：0
页码：134-140
语言：中文
分类：Q51[生物学—生物化学] S511.01[农业科学—作物学]
作者机构：[1]State Key Laboratory of Proteomics, Beijing Proteome Research Center, Beijing Institute of Radiation Medicine, Beijing 102206, China, [2]Institutes of Biomedical Sciences, Fudan University, Shanghai 200032, China
相关基金：Funding This work was supported by grants from the National HighTec Research Developing Programme （2006AA02A310） of China, the Special Funds for Major State Basic Research of China （2006CB910802）, the National Natural Science Foundation of China for Innovation group （30621063）, and the State Key Laboratory of Proteomics （SKLP-K200801）.
相关项目：功能化开管毛细管色谱柱制备及其在复杂生物样本的磷酸化蛋白质分析中的应用研究

关键词：串联亲和纯化, 蛋白复合物, 鉴定, TSC 1, tandem affinity purification, proteomics

中文摘要：

在 TSC1 和 TSC2 基因的变化导致有块茎的硬化建筑群(TSC ) ，它被影响多重纸巾的智力迟钝，癫痫，和良性的肿瘤临床上描绘。TSC 蛋白质建筑群的众多的部件仍然是 uncharacterized。这里，我们用 proteomic 策略在生理的条件下面报导 TSC1 建筑群的纯化。我们用一个双人脚踏车亲密关系纯化方法净化了 TSC1 蛋白质建筑群并且识别了包含 139 个部件的蛋白质建筑群。TSC1 (TSC2 和 DOCK7 ) 的二已知的有约束力的蛋白质与另外的新潜在的搭挡一起被识别，盖子报导了它并且新奇 TSC1 功能的范畴。生物信息学和生物化学的方法被用来评估观察 proteinprotein 相互作用。有 TSC1 的出版表示 proteomics/genomics 研究的比较分析揭示了可能机能上地相关的超过 20 个普通候选人。数据集合提供扩展我们 TSC1 的函数和 TSC 的机制的知识的新方向。结果是高度可靠的，它被 TSC1 和许多 TSC1/2-regulated 蛋白质的一些报导搭挡的鉴定反映。有趣地，许多新功能的范畴被识别，例如 DNA 修理，它提供新奇提示给 TSC1 的功能。而且，一些 neuronal 可能调整神经原的正常功能的疾病相关的蛋白质被识别。因此，结果新相互作用建议那许多应该是生物学上意义。进一步调查这些部件的规章的机制将有趣。

英文摘要：

Mutations in the TSC1 and TSC2 genes lead to tuberous sclerosis complex （TSC）, which is characterized clinically by mental retardation, epilepsy, and benign tumors affect- ing multiple tissues. Numerous components of the TSC protein complex remain uncharacterized. Here we report the purification of the TSCI complex under physiological conditions using a proteomic strategy. We purified the TSC1 protein complex using a tandem affinity purifi- cation method and identified a protein complex contain- ing 139 components. Two known binding proteins of TSC1 （TSC2 and DOCK7） were identified along with other new potential partners, which cover reported and novel TSC1 functional categories. Bioinformatics and bio- chemical methods were used to evaluate the observed protein-protein interactions. A comparative analysis with a published expression proteomics/genomics study of TSC1 revealed more than 20 common candidates that might be functionally relevant. The data set provides new directions in which to expand our knowledge of the func- tions of TSC1 and the mechanisms of TSC. The results are highly reliable, which is reflected by the identification of a few reported partners of TSC1 and many TSCI/2- regulated proteins. Interestingly, many new functional categories were identified, such as DNA repair, which provide novel hints to the function of TSC1. Moreover, a few neuronal disease-related proteins that might regulate the normal functions of neurons were identified. Thus, the results suggest that many of the new interactions should be biologically significance. It will be interesting to further investigate the regulatory mechanisms of these components.