In addition to the highly targeted quantification of metabolites already known to play major roles in oxidative stress, to provide data directly compatible with current models, we will also take an untargeted metabolomics approach. This will enable us to identify other areas of the metabolome influenced by, or influencing, oxidative stress and will allow us to compare changes in each of the stress-inducing stimuli. We have recently pioneered untargeted metabolite profiling of T. brucei using ultra-high resolution mass spectrometry (Kamleh et al. 2008 PMID: 18470888). This will link directly to the modelling in WP1 by providing the high-accuracy metabolome data for the ab inito networks. We have, so far, been able to identify in the order of 400 metabolites in trypanosomes using a single, simple sample preparation protocol. The untargeted metabolite approach enables perturbations to multiple parts of the cellular metabolome to be assessed in a single experiment and also point to changes in metabolism secondary to inhibition of target enzymes. For example, changes to NADP/NADPH balance can affect other biochemical pathways i.e. secondary effects will ripple out through the metabolome as a consequence of primary changes. Data obtained on changes to metabolite levels will be compared to dynamic changes in transcript and protein levels. In this manner we hope to extend our knowledge of pathways involved in oxidative stress defence, beyond the already established pathways involved in trypanothione production.
SEEK ID: https://fairdomhub.org/studies/103
Experimentalists: Dong-Hyun Kim
Created: 10th Dec 2012 at 17:10