Comparative proteomics profiling of a gentamicin-attenuated Leishmania infantum cell line identifies key changes in parasite thiol-redox metabolism

Hamid Daneshvar, Susan Wyllie, Stephen Phillips, Paul Hagan, Richard Burchmore

    Research output: Contribution to journalArticle

    32 Citations (Scopus)

    Abstract

    We have previously described an attenuated line of Leishmania infantum (H-line), selected by culturing promastigotes in vitro in the presence of gentamicin. To elucidate the molecular basis for this attenuation, we undertook a comparative proteomic analysis using multiplex 2-dimensional (2D) difference gel electrophoresis. Eighteen proteins that showed significant and reproducible changes in expression were identified. Many of these were components of the thiol-redox control system in Leishmania and this observation, validated by Western blot, prompted us to investigate the sensitivity of the attenuated line to oxidative stress. The attenuated line was found to be significantly more susceptible to hydrogen peroxide, a change which may explain the loss of virulence. In a direct assay of trypanothione-dependent peroxidase activity, hydrogen peroxide metabolism in the H-line was significantly lower than in wild type. Furthermore, trypanothione reductase activity was significantly lower in the H-line, suggesting that gentamicin selection may result in pleiotropic affects on thiol metabolism in Leishmania. A putative RNA-binding protein was very strongly up-regulated in the attenuated line, suggesting a possible target for gentamicin in Leishmania.
    Original languageEnglish
    Pages (from-to)1463-1471
    Number of pages9
    JournalJournal of Proteomics
    Volume75
    Issue number5
    Early online date28 Nov 2011
    DOIs
    Publication statusPublished - 16 Feb 2012

    Keywords

    • Leishmania
    • Attenuation
    • Virulence
    • Vaccine

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