Abstract
Mass spectrometry (MS) offers a sensitive, reliable, and highly accurate method for measurement of isotopic labeling, which is required for generating comprehensive flux maps using metabolic flux analysis (MFA). We present protocols for assessing isotope labeling in a wide range of biochemical species, including proteinogenic amino acids, free organic and amino acids, sugar phosphates, lipids, starch-glucose, and RNA-ribose. We describe the steps of sample preparation, MS analysis, and data handling required to obtain high-quality isotope labeling measurements that are applicable to MFA. By selecting target analytes that maximize identifiability of the key fluxes of interest, MS measurements of isotope labeling can provide a powerful platform for assessing metabolic fluxes in complex biochemical networks.
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Acknowledgments
This work has been supported by NSF CAREER Award CBET-0955251 (to J.D.Y).
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Young, J.D., Allen, D.K., Morgan, J.A. (2014). Isotopomer Measurement Techniques in Metabolic Flux Analysis II: Mass Spectrometry. In: Sriram, G. (eds) Plant Metabolism. Methods in Molecular Biology, vol 1083. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-62703-661-0_7
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DOI: https://doi.org/10.1007/978-1-62703-661-0_7
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