Proteomic analyses of murine macrophages treated with Bacillus anthracis lethal toxin

R. Sapra, S. P. Gaucher, J. S. Lachmann, G. M. Buffleben, G. S. Chirica, J. E. Comer, J. W. Peterson, A. K. Chopra, A. K. Singh

Research output: Contribution to journalArticlepeer-review

28 Scopus citations


Bacillus anthracis is the etiological agent of anthrax and the bacterium produces a tripartite anthrax toxin composed of protective antigen (PA), lethal factor (LF) and edema factor (EF). PA represents the binding domain of the toxin and acts in concert with either LF, a metalloprotease, or EF, an adenylate cyclase, to form lethal toxin (LeTx) or edema toxin (EdTx), respectively. We analyzed the proteomics response of two murine macrophage cell lines (J774.1A and RAW264.7) following B. anthracis LeTx treatment to detect unique host proteins involved in anthrax infection using difference in-gel electrophoresis (DIGE) followed by nanoLC-MS for identification of the proteins. The comparative proteomics approach identified a set of proteins in each cell line that was consistently upregulated when the two macrophage cell lines were treated with LeTx. The upregulated proteins include those involved in energy metabolism, cytoskeleton structure and stress response. A subset of five proteins (ATP synthase β subunit, β-actin, Hsp70, vimentin, and Hsp60 homolog) was identified that were commonly upregulated in both cell lines. The proteomic data suggest the involvement of reactive oxygen species (ROS) in cell lysis as seen by the upregulation of proteins that lead to the production of ROS in both the cell lines used in our study. However, proteins that afford protection against ROS may play an important role in the survival of the macrophage to LeTx infection as shown by the differences in proteomic responses of the two cell lines to the action of LeTx. These identified proteins may have the potential to be used as biomarkers for diagnostics and therapeutics.

Original languageEnglish (US)
Pages (from-to)157-167
Number of pages11
JournalMicrobial Pathogenesis
Issue number4-5
StatePublished - Oct 2006


  • Bacillus anthracis
  • Difference in-gel electrophoresis (DIGE)
  • Lethal toxin
  • Macrophage cell lines
  • Reactive oxygen species (ROS)

ASJC Scopus subject areas

  • Microbiology
  • Infectious Diseases


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