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Analysis of Kinetic Labeling of Amino Acids and Organic Acids by GC-MS

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http://pubman.mpdl.mpg.de/cone/persons/resource/persons97052

Araujo,  W. L.
Central Metabolism, Department Willmitzer, Max Planck Institute of Molecular Plant Physiology, Max Planck Society;

http://pubman.mpdl.mpg.de/cone/persons/resource/persons97445

Tohge,  T.
Central Metabolism, Department Willmitzer, Max Planck Institute of Molecular Plant Physiology, Max Planck Society;

http://pubman.mpdl.mpg.de/cone/persons/resource/persons97323

Nunes-Nesi,  A.
Central Metabolism, Department Willmitzer, Max Planck Institute of Molecular Plant Physiology, Max Planck Society;

http://pubman.mpdl.mpg.de/cone/persons/resource/persons97324

Obata,  T.
Central Metabolism, Department Willmitzer, Max Planck Institute of Molecular Plant Physiology, Max Planck Society;

http://pubman.mpdl.mpg.de/cone/persons/resource/persons97147

Fernie,  A. R.
Central Metabolism, Department Willmitzer, Max Planck Institute of Molecular Plant Physiology, Max Planck Society;

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Citation

Araujo, W. L., Tohge, T., Nunes-Nesi, A., Obata, T., & Fernie, A. R. (2014). Analysis of Kinetic Labeling of Amino Acids and Organic Acids by GC-MS. Methods in molecular biology, 1090, 107-119. doi:10.1007/978-1-62703-688-7_7.


Cite as: http://hdl.handle.net/11858/00-001M-0000-0014-B3B1-F
Abstract
Plant metabolic pathways and the molecular and atomic fluxes through them can be deduced using stable isotopically labeled substrates. To this end one prerequisite is accurate measurement of the labeling pattern of targeted metabolites. Experiments are generally limited to the use of single-element isotopes, mainly (13)C. Here, we summarize the application of gas chromatography-time of flight mass spectrometry (GC-TOF-MS) for metabolic studies using differently labeled elemental isotopes applied to both intact organelles and whole plant tissue. This method allows quantitative evaluation of a broad range of metabolic pathways without the need for laborious (and potentially inaccurate) chemical fractionation procedures commonly used in the estimation of fluxes following incubation in radiolabeled substrates. We focus herein on the determination of isotope labeling in organic and amino acids.