Comparative proteomics of human monkeypox and vaccinia intracellular mature and extracellular enveloped virions

Nathan P. Manes, Ryan Estep, Heather M. Mottaz, Ronald J. Moore, Therese R W Clauss, Matthew E. Monroe, Xiuxia Du, Joshua N. Adkins, Scott Wong, Richard D. Smith

Research output: Contribution to journalArticle

41 Citations (Scopus)

Abstract

Orthopoxviruses are among the largest and most complex of the animal viruses. In response to the recent emergence of monkeypox in Africa and the threat of smallpox bioterrorism, two orthopoxviruses with different pathogenic potentials, human monkeypox virus and vaccinia virus, were proteomically compared with the goal of identifying proteins required for pathogenesis. Orthopoxviruses were grown in HeLa cells to two different viral forms (intracellular mature virus and extracellular enveloped virus), purified by sucrose gradient ultracentrifugation, denatured using RapiGest surfactant, and digested with trypsin. Unfractionated samples and strong cation exchange HPLC fractions were analyzed by high-resolution reversed-phase nano-LC-MS/MS, and analyses of the MS/MS spectra using SEQUEST and X! Tandem resulted in the confident identification of hundreds of monkeypox, vaccinia, and copurified host-cell proteins. The unfractionated samples were additionally analyzed by LC-MS using an LTQ-Orbitrap, and the accurate mass and elution time tag approach was used to perform quantitative comparisons. Possible pathophysiological roles of differentially abundant Orthopoxvirus proteins are discussed. Data, processed results, and protocols are available at http://www.proteomicsresource. org/.

Original languageEnglish (US)
Pages (from-to)960-968
Number of pages9
JournalJournal of Proteome Research
Volume7
Issue number3
DOIs
StatePublished - Mar 2008

Fingerprint

Monkeypox
Orthopoxvirus
Vaccinia
Viruses
Virion
Proteomics
Monkeypox virus
Bioterrorism
Proteins
Smallpox
Vaccinia virus
Ultracentrifugation
HeLa Cells
Surface-Active Agents
Trypsin
Sucrose
Cations
High Pressure Liquid Chromatography
Animals

Keywords

  • Mass spectrometry
  • Monkeypox
  • Proteomics
  • SEQUEST
  • Vaccinia
  • XI tandem

ASJC Scopus subject areas

  • Biochemistry
  • Chemistry(all)
  • Genetics
  • Biotechnology

Cite this

Manes, N. P., Estep, R., Mottaz, H. M., Moore, R. J., Clauss, T. R. W., Monroe, M. E., ... Smith, R. D. (2008). Comparative proteomics of human monkeypox and vaccinia intracellular mature and extracellular enveloped virions. Journal of Proteome Research, 7(3), 960-968. https://doi.org/10.1021/pr070432+

Comparative proteomics of human monkeypox and vaccinia intracellular mature and extracellular enveloped virions. / Manes, Nathan P.; Estep, Ryan; Mottaz, Heather M.; Moore, Ronald J.; Clauss, Therese R W; Monroe, Matthew E.; Du, Xiuxia; Adkins, Joshua N.; Wong, Scott; Smith, Richard D.

In: Journal of Proteome Research, Vol. 7, No. 3, 03.2008, p. 960-968.

Research output: Contribution to journalArticle

Manes, NP, Estep, R, Mottaz, HM, Moore, RJ, Clauss, TRW, Monroe, ME, Du, X, Adkins, JN, Wong, S & Smith, RD 2008, 'Comparative proteomics of human monkeypox and vaccinia intracellular mature and extracellular enveloped virions', Journal of Proteome Research, vol. 7, no. 3, pp. 960-968. https://doi.org/10.1021/pr070432+
Manes, Nathan P. ; Estep, Ryan ; Mottaz, Heather M. ; Moore, Ronald J. ; Clauss, Therese R W ; Monroe, Matthew E. ; Du, Xiuxia ; Adkins, Joshua N. ; Wong, Scott ; Smith, Richard D. / Comparative proteomics of human monkeypox and vaccinia intracellular mature and extracellular enveloped virions. In: Journal of Proteome Research. 2008 ; Vol. 7, No. 3. pp. 960-968.
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