Isotopomer-based metabolomic analysis by NMR and mass spectrometry.
Nuclear magnetic resonance (NMR) and mass spectrometry (MS) together are synergistic in their ability to profile comprehensively the metabolome of cells and tissues. In addition to identification and quantification of metabolites, changes in metabolic pathways and fluxes in response to external perturbations can be reliably determined by using stable isotope tracer methodologies. NMR and MS together are able to define both positional isotopomer distribution in product metabolites that derive from a given stable isotope-labeled precursor molecule and the degree of enrichment at each site with good precision. Together with modeling tools, this information provides a rich functional biochemical readout of cellular activity and how it responds to external influences. In this chapter, we describe NMR- and MS-based methodologies for isotopomer analysis in metabolomics and its applications for different biological systems.
Lane
Andrew N
AN
JG Brown Cancer Center, University of Louisville, Louisville, Kentucky 40202, USA.
Fan
Teresa W-M
TW
Higashi
Richard M
RM
D000818:Animals; D001383:Azetidinecarboxylic Acid / Q000031:analogs & derivatives / Q000737:chemistry; D002457:Cell Extracts; D045744:Cell Line, Tumor; D006801:Humans; D007553:Isotope Labeling; D007554:Isotopes; D009682:Magnetic Resonance Spectroscopy / Q000379:methods*; D013058:Mass Spectrometry / Q000379:methods*; D051379:Mice; D041681:NIH 3T3 Cells; D011522:Protons; D012015:Reference Standards
D002457:Cell Extracts; D007554:Isotopes; D011522:Protons; D001383:Azetidinecarboxylic Acid; C052863:mugineic acid