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Submitted on April 14, 2006
Accepted on May 8, 2006

Analysis of the salmonella typhimurium proteome through environmental response towards infectious conditions

Joshua N. Adkins, Heather M. Mottaz, Angela D. Norbeck, Jean K. Gustin, Joanne Rue, Therese R.W. Clauss, Samuel O. Purvine, Karin Rodland, Fred Heffron, and Richard D. Smith

MACROMOLECULAR STRUCTURE & DYNAMICS, Pacific Northwest National Laboratory, Richland, Washington 99352

Corresponding Author: rds{at}pnl.gov

Salmonella enterica serovar Typhimurium (aka, S. typhimurium) is a facultative intracellular pathogen that causes ~40,000 reported cases of acute gastroenteritis and diarrhea each year in the United States. While many successful physiological, biochemical, and genetic approaches have been taken to determine the key virulence determinants encoded by this organism, the shear number of uncharacterized reading frames observed within the S. enterica genome suggests that many more virulence factors remain to be discovered. We have used a liquid chromatography-mass spectrometry-based “bottom-up” proteomics approach to generate a more complete picture of the gene products that S. typhimurium synthesizes under typical laboratory conditions, as well as in culture media that are known to induce expression of virulence genes. When grown to logarithmic (log) phase in rich medium, S. typhimurium is known to express many genes that are required for invasion of epithelial cells. Conversely, stationary phase cultures of S. typhimurium express genes that are needed for both systemic infection and growth within infected macrophages. Lastly, bacteria grown in an acidic, magnesium-depleted minimal medium (MgM) designed to mimic the phagocytic vacuole have been shown to upregulate virulence gene expression. Initial comparisons of protein abundances from bacteria grown under each of these conditions indicated that the majority of proteins do not change significantly. However, we observed subsets of proteins whose expression was largely restricted to one of the 3 culture conditions. For example, cells grown in MgM had a higher abundance of Mg2+ transport proteins than found in other growth conditions. A second more virulent S. typhimurium strain (14028) was also cultured under these same growth conditions, and the results were directly compared to those obtained for strain LT2. This comparison offered a unique opportunity to contrast protein populations in these closely related bacteria.


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