Journal of Biomolecular NMR

, Volume 49, Issue 3–4, pp 267–280 | Cite as

NMR-based stable isotope resolved metabolomics in systems biochemistry

Article

Abstract

An important goal of metabolomics is to characterize the changes in metabolic networks in cells or various tissues of an organism in response to external perturbations or pathologies. The profiling of metabolites and their steady state concentrations does not directly provide information regarding the architecture and fluxes through metabolic networks. This requires tracer approaches. NMR is especially powerful as it can be used not only to identify and quantify metabolites in an unfractionated mixture such as biofluids or crude cell/tissue extracts, but also determine the positional isotopomer distributions of metabolites derived from a precursor enriched in stable isotopes such as 13C and 15N via metabolic transformations. In this article we demonstrate the application of a variety of 2-D NMR editing experiments to define the positional isotopomers of compounds present in polar and non-polar extracts of human lung cancer cells grown in either [U–13C]-glucose or [U–13C,15N]-glutamine as source tracers. The information provided by such experiments enabled unambiguous reconstruction of metabolic pathways, which is the foundation for further metabolic flux modeling.

Keywords

Stable isotope resolved metabolomics Isotope editing 

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Copyright information

© Springer Science+Business Media B.V. 2011

Authors and Affiliations

  1. 1.Department of ChemistryUniversity of LouisvilleLouisvilleUSA
  2. 2.Center for Regulatory Environmental Analytical MetabolomicsUniversity of LouisvilleLouisvilleUSA
  3. 3.JG Brown Cancer CenterUniversity of LouisvilleLouisvilleUSA
  4. 4.LouisvilleUSA

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