Proteomic analysis of SARS associated coronavirus using two-dimensional liquid chromatography mass spectrometry and one-dimensional sodium dodecyl sulfate-polyacrylamide gel electrophoresis followed by mass spectroemtric analysis
Proteomic analysis of SARS associated coronavirus using two-dimensional liquid chromatography mass spectrometry and one-dimensional sodium dodecyl sulfate-polyacrylamide gel electrophoresis followed by mass spectroemtric analysis
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DOI：
10.1021/pr034111j
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发表时间：
2004-05-01
影响因子：
4.4
通讯作者：
Wu, JR
中科院分区：
文献类型：
--
作者：
Zeng, R;Ruan, HQ;Wu, JR
本研究检测了严重急性呼吸综合征相关冠状病毒（SARS-CoV）及其感染的 Vero E6 细胞的蛋白质组。通过一维电泳随后 ESI-MS/MS 鉴定或通过二维液相色谱-ESI-MS/MS 鸟枪策略分析病毒感染细胞的胞质和细胞核部分以及粗制病毒粒子。首次鉴定了 SARS-CoV 的所有四种预测结构蛋白，包括 S（刺突）、M（膜）、N（核衣壳）和 E（包膜）蛋白。此外，还观察到 M 蛋白的一个新的磷酸化位点。这些凝胶方法和非凝胶方法的结合为 SARS-CoV 蛋白质组提供了快速且互补的方法，可广泛用于其他病毒的分析。
The proteomes of the severe acute respiratory syndrome-associated coronavirus (SARS-CoV) and its infected Vero E6 cells were detected in the present study. The cytosol and nucleus fractions of virus infected cells as well as the crude virions were analyzed either by one-dimensional electrophoresis followed by ESI-MS/MS identification or by shotgun strategy with two-dimensional liquid chromatography-ESI-MS/MS. For the first time, all of the four predicted structural proteins of SARS-CoV were identified, including S (Spike), M (Membrane), N (Nucleocapsid), and E (Envolope) proteins. In addition, a novel phosphorylated site of M protein was observed. The combination of these gel-base and non-gel methods provides fast and complimentary approaches to SARS-CoV proteome and can be widely used in the analysis of other viruses.