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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Authors: | Zeng Rong Ruan Hong-Qiang Jiang Xiao-Sheng Zhou Hu Shi Lv Zhang Lei Sheng Quan-Hu Tu Qiang Xia Qi-Chang Wu Jia-Rui |
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Institution: | Research Center for Proteome Analysis, Key Lab of Proteomics, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, 320 YueYang Road, Shanghai 200031, China. zr@sibs.ac.cn |
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Abstract: | 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. |
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