NMR-based stable isotope resolved metabolomics in systems biochemistry
- 26 February 2011
- journal article
- research article
- Published by Springer Nature in Journal of Biomolecular NMR
- Vol. 49 (3-4), 267-280
- https://doi.org/10.1007/s10858-011-9484-6
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
This publication has 41 references indexed in Scilit:
- Stable isotope-resolved metabolomic analysis of lithium effects on glial-neuronal metabolism and interactionsMetabolomics, 2010
- Isotopomer analysis of lipid biosynthesis by high resolution mass spectrometry and NMRAnalytica Chimica Acta, 2009
- Altered regulation of metabolic pathways in human lung cancer discerned by 13C stable isotope-resolved metabolomics (SIRM)Molecular Cancer, 2009
- Rhabdomyosarcoma cells show an energy producing anabolic metabolic phenotype compared with primary myocytesMolecular Cancer, 2008
- Beyond aerobic glycolysis: Transformed cells can engage in glutamine metabolism that exceeds the requirement for protein and nucleotide synthesisProceedings of the National Academy of Sciences, 2007
- Quantification of Intracellular Phosphorylated Carbohydrates in HT29 Human Colon Adenocarcinoma Cell Line Using Liquid Chromatography−Electrospray Ionization Tandem Mass SpectrometryAnalytical Chemistry, 2007
- Mammalian Pyrimidine Biosynthesis: Fresh Insights into an Ancient PathwayJournal of Biological Chemistry, 2004
- Heteronuclear Scalar Couplings in the Bases and Sugar Rings of Nucleic Acids: Their Determination and Application in Assignment and Conformational AnalysisMagnetic Resonance in Chemistry, 1996
- Metabolite profiling by one- and two-dimensional NMR analysis of complex mixturesProgress in Nuclear Magnetic Resonance Spectroscopy, 1996
- Glucose and glutamine metabolism in C6 glioma cells studied by carbon 13 NMRBiochimie, 1996