Global analysis of the Deinococcus radiodurans proteome by using accurate mass tags
Global analysis of the Deinococcus radiodurans proteome by using accurate mass tags
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DOI：
10.1073/pnas.172170199
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发表时间：
2002-08-20
影响因子：
11.1
通讯作者：
Smith, RD
中科院分区：
文献类型：
--
作者：
Lipton, MS;Pasa-Tolic, L;Smith, RD
了解生物系统及其组成部分的作用是通过定量，灵敏和全面测量其蛋白质组如何变化的能力来促进的，例如，以应对环境的扰动。为此，我们已经开发出一种高通量的方法来表征生物体的动态蛋白质组的基础上相结合的全球酶消化，高分辨率液相色谱分离，傅立叶变换离子回旋共振质谱分析。所产生的肽作为蛋白质的精确质量标签，并已用于鉴定电离辐射抗性细菌耐辐射异常球菌的预测蛋白质组，置信度>61%。这一部分代表了迄今为止任何生物体最广泛的蛋白质组覆盖范围，包括715种以前被注释为假设或保守假设的蛋白质。
Understanding biological systems and the roles of their constituents is facilitated by the ability to make quantitative, sensitive, and comprehensive measurements of how their proteome changes, e.g., in response to environmental perturbations. To this end, we have developed a high-throughput methodology to characterize an organism's dynamic proteome based on the combination of global enzymatic digestion, high-resolution liquid chromatographic separations, and analysis by Fourier transform ion cyclotron resonance mass spectrometry. The peptides produced serve as accurate mass tags for the proteins and have been used to identify with high confidence >61% of the predicted proteome for the ionizing radiation-resistant bacterium Deinococcus radiodurans. This fraction represents the broadest proteome coverage for any organism to date and includes 715 proteins previously annotated as either hypothetical or conserved hypothetical.