Performing comparative peptidomics analyses of Salmonella from different growth conditions.

Joshua N. Adkins, Heather Mottaz, Thomas O. Metz, Charles Ansong, Nathan P. Manes, Richard D. Smith, Fred Heffron

    Research output: Contribution to journalArticle

    2 Scopus citations

    Abstract

    Host-pathogen interactions are complex competitions during which both the host and the pathogen adapt rapidly to each other in order for one or the other to survive. Salmonella enterica serovar Typhimurium is a pathogen with a broad host range that causes a typhoid fever-like disease in mice and severe food poisoning in humans. The murine typhoid fever is a systemic infection in which S. typhimurium evades part of the immune system by replicating inside macrophages and other cells. The transition from a foodborne contaminant to an intracellular pathogen must occur rapidly in multiple, ordered steps in order for S. typhimurium to thrive within its host environment. Using S. typhimurium isolated from rich culture conditions and from conditions that mimic the hostile intracellular environment of the host cell, a native low molecular weight protein fraction, or peptidome, was enriched from cell lysates by precipitation of intact proteins with organic solvents. The enriched peptidome was analyzed by both LC-MS/MS and LC-MS-based methods, although several other methods are possible. Pre-fractionation of peptides allowed identification of small proteins and protein degradation products that would normally be overlooked. Comparison of peptides present in lysates prepared from Salmonella grown under different conditions provided a unique insight into cellular degradation processes as well as identification of novel peptides encoded in the genome but not annotated. The overall approach is detailed here as applied to Salmonella and is adaptable to a broad range of biological systems.

    Original languageEnglish (US)
    Pages (from-to)13-27
    Number of pages15
    JournalMethods in molecular biology (Clifton, N.J.)
    Volume615
    DOIs
    StatePublished - 2010

    ASJC Scopus subject areas

    • Molecular Biology
    • Genetics

    Fingerprint Dive into the research topics of 'Performing comparative peptidomics analyses of Salmonella from different growth conditions.'. Together they form a unique fingerprint.

  • Cite this