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.
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.
Zeng
Rong
R
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
Ruan
Hong-Qiang
HQ
Jiang
Xiao-Sheng
XS
Zhou
Hu
H
Shi
Lv
L
Zhang
Lei
L
Sheng
Quan-Hu
QH
Tu
Qiang
Q
Xia
Qi-Chang
QC
Wu
Jia-Rui
JR
D000595:Amino Acid Sequence; D002853:Chromatography, Liquid; D004591:Electrophoresis, Polyacrylamide Gel; D013058:Mass Spectrometry; D008969:Molecular Sequence Data; D019251:Nucleocapsid / Q000378:metabolism*; D010455:Peptides / Q000378:metabolism*; D010766:Phosphorylation; D045473:Severe acute respiratory syndrome-related coronavirus / Q000378:metabolism*; D014759:Viral Envelope Proteins / Q000378:metabolism*; D014763:Viral Matrix Proteins / Q000378:metabolism*
D010455:Peptides; D014759:Viral Envelope Proteins; D014763:Viral Matrix Proteins