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.
|Number of pages||14|
|Journal||Journal of Biomolecular NMR|
|State||Published - Apr 2011|
Bibliographical noteFunding Information:
Acknowledgments This work was supported in part by National Science Foundation EPSCoR grant # EPS-0447479; National Institutes of Health Grant numbers 5P20RR018733, 1R01CA118434-01A2, 3R01CA118434-02S1, R21CA133668-01; the Kentucky Challenge for Excellence, and the Brown Foundation. We thank Ms. JinLian Tan for expert technical assistance, and Dr. S. Arumugam for assistance with NMR experiments.
- Isotope editing
- Stable isotope resolved metabolomics
ASJC Scopus subject areas