A high-temporal resolution technology for dynamic proteomic analysis based on 35S labeling.

A high-temporal resolution technology for dynamic proteomic analysis based on 35S labeling.
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基于35S标记的动态蛋白质组分析的高时间分辨率技术。

DOI:
10.1371/journal.pone.0002991
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发表时间:
2008-08-20
期刊:
影响因子:
3.7
通讯作者:
He D
He D
中科院分区:
综合性期刊3区
文献类型:
--
作者:
Zhang Z;Chen J;Guo F;He L;Wu Y;Zeng C;Xiao X;He D

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随着越来越多的研究工作被动态或差异蛋白质组学所吸引,需要一种具有高时间分辨率和高通量的方法。在本研究中,通过分析高度同步的A549细胞以及大鼠肝脏2/3部分肝切除手术中的动态蛋白质组变化,设计并测试了动态蛋白质组学的35S体内标记分析(SiLAD)。结果验证了SiLAD技术与二维电泳相结合,提供了一种高灵敏度的方法来说明不受干扰的内源蛋白动态变化,具有良好的时间分辨率和高信噪比。通过这种技术可以发现或重新分类大量的差异蛋白。 SiLAD的另一个独特功能是能够量化蛋白质表达率,从而更有效地反映细胞生理转折点。最后,规定的 SiLAD 蛋白质组快照模式有可能用作表征良好调节的生物过程中每个阶段的专有符号。
As more and more research efforts have been attracted to dynamic or differential proteomics, a method with high temporal resolution and high throughput is required. In present study, a 35S in vivo Labeling Analysis for Dynamic Proteomics (SiLAD) was designed and tested by analyzing the dynamic proteome changes in the highly synchronized A549 cells, as well as in the rat liver 2/3 partial hepatectomy surgery. The results validated that SiLAD technique, in combination with 2-Dimensional Electrophoresis, provided a highly sensitivity method to illustrate the non-disturbed endogenous proteins dynamic changes with a good temporal resolution and high signal/noise ratio. A significant number of differential proteins can be discovered or re-categorized by this technique. Another unique feature of SiLAD is its capability of quantifying the rate of protein expression, which reflects the cellular physiological turn points more effectively. Finally, the prescribed SiLAD proteome snapshot pattern could be potentially used as an exclusive symbol for characterizing each stage in well regulated biological processes.
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