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

J Proteome Res. 2004 May-Jun;3(3):549-55. doi: 10.1021/pr034111j.

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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Chromatography, Liquid
  • Electrophoresis, Polyacrylamide Gel
  • Mass Spectrometry
  • Molecular Sequence Data
  • Nucleocapsid / metabolism*
  • Peptides / metabolism*
  • Phosphorylation
  • Severe acute respiratory syndrome-related coronavirus / metabolism*
  • Viral Envelope Proteins / metabolism*
  • Viral Matrix Proteins / metabolism*

Substances

  • Peptides
  • Viral Envelope Proteins
  • Viral Matrix Proteins