title: Pox proteomics: mass spectrometry analysis and identification of Vaccinia virion proteins.
abstract: BACKGROUND
  Although many vaccinia virus proteins have been identified and studied in detail, only a few studies have attempted a comprehensive survey of the protein composition of the vaccinia virion. These projects have identified the major proteins of the vaccinia virion, but little has been accomplished to identify the unknown or less abundant proteins. Obtaining a detailed knowledge of the viral proteome of vaccinia virus will be important for advancing our understanding of orthopoxvirus biology, and should facilitate the development of effective antiviral drugs and formulation of vaccines.
  
  
  RESULTS
  In order to accomplish this task, purified vaccinia virions were fractionated into a soluble protein enriched fraction (membrane proteins and lateral bodies) and an insoluble protein enriched fraction (virion cores). Each of these fractions was subjected to further fractionation by either sodium dodecyl sulfate-polyacrylamide gel electophoresis, or by reverse phase high performance liquid chromatography. The soluble and insoluble fractions were also analyzed directly with no further separation. The samples were prepared for mass spectrometry analysis by digestion with trypsin. Tryptic digests were analyzed by using either a matrix assisted laser desorption ionization time of flight tandem mass spectrometer, a quadrupole ion trap mass spectrometer, or a quadrupole-time of flight mass spectrometer (the latter two instruments were equipped with electrospray ionization sources). Proteins were identified by searching uninterpreted tandem mass spectra against a vaccinia virus protein database created by our lab and a non-redundant protein database.
  
  
  CONCLUSIONS
  Sixty three vaccinia proteins were identified in the virion particle. The total number of peptides found for each protein ranged from 1 to 62, and the sequence coverage of the proteins ranged from 8.2% to 94.9%. Interestingly, two vaccinia open reading frames were confirmed as being expressed as novel proteins: E6R and L3L.
authors.affiliation: Oregon State University, Department of Microbiology, 220 Nash Hall, Corvallis, OR 97331-3804, USA. yoderj@science.oregonstate.edu
authors.forename: Jennifer D
authors.initials: JD
authors.lastname: Yoder
authors.forename: Tsefang S
authors.initials: TS
authors.lastname: Chen
authors.forename: Cliff R
authors.initials: CR
authors.lastname: Gagnier
authors.forename: Srilakshmi
authors.initials: S
authors.lastname: Vemulapalli
authors.forename: Claudia S
authors.initials: CS
authors.lastname: Maier
authors.forename: Dennis E
authors.initials: DE
authors.lastname: Hruby
mesh_terms: D000595:Amino Acid Sequence; D030562:Databases, Protein; D004591:Electrophoresis, Polyacrylamide Gel; D013058:Mass Spectrometry / Q000379:methods*; D008969:Molecular Sequence Data; D016366:Open Reading Frames / Q000235:genetics; D010455:Peptides / Q000737:chemistry; D020543:Proteome; D040901:Proteomics; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D014357:Trypsin / Q000378:metabolism; D014616:Vaccinia virus / Q000254:growth & development / Q000378:metabolism*; D014764:Viral Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D014771:Virion / Q000378:metabolism*
chemical_list: D010455:Peptides; D020543:Proteome; D014764:Viral Proteins; D014357:Trypsin