title: Stable isotope-assisted metabolomics for network-wide metabolic pathway elucidation.
abstract: The combination of high-resolution LC-MS-based untargeted metabolomics with stable isotope tracing provides a global overview of the cellular fate of precursor metabolites. This methodology enables detection of putative metabolites from biological samples and simultaneous quantification of the pattern and extent of isotope labeling. Labeling of Trypanosoma brucei cell cultures with 50% uniformly (13)C-labeled glucose demonstrated incorporation of glucose-derived carbon into 187 of 588 putatively identified metabolites in diverse pathways including carbohydrate, nucleotide, lipid, and amino acid metabolism. Labeling patterns confirmed the metabolic pathways responsible for the biosynthesis of many detected metabolites, and labeling was detected in unexpected metabolites, including two higher sugar phosphates annotated as octulose phosphate and nonulose phosphate. This untargeted approach to stable isotope tracing facilitates the biochemical analysis of known pathways and yields rapid identification of previously unexplored areas of metabolism.
authors.affiliation: Wellcome Trust Centre for Molecular Parasitology, Institute of Infection, Immunity and Inflammation, College of Medical, Veterinary and Life Sciences, University of Glasgow, Glasgow, G12 8TA, UK.
authors.forename: Darren J
authors.initials: DJ
authors.lastname: Creek
authors.forename: Achuthanunni
authors.initials: A
authors.lastname: Chokkathukalam
authors.forename: Andris
authors.initials: A
authors.lastname: Jankevics
authors.forename: Karl E V
authors.initials: KE
authors.lastname: Burgess
authors.forename: Rainer
authors.initials: R
authors.lastname: Breitling
authors.forename: Michael P
authors.initials: MP
authors.lastname: Barrett
mesh_terms: D002247:Carbon Isotopes / Q000032:analysis / Q000378:metabolism; D002853:Chromatography, Liquid / Q000191:economics / Q000379:methods; D005947:Glucose / Q000032:analysis / Q000378:metabolism; D007553:Isotope Labeling / Q000191:economics / Q000379:methods; D013058:Mass Spectrometry / Q000191:economics / Q000379:methods; D053858:Metabolic Networks and Pathways; D055432:Metabolomics / Q000191:economics / Q000379:methods*; D013997:Time Factors; D014346:Trypanosoma brucei brucei / Q000378:metabolism*; D014353:Trypanosomiasis, African / Q000469:parasitology
chemical_list: D002247:Carbon Isotopes; D005947:Glucose