Analysis of fatty acid metabolism using stable isotope tracers and mass spectrometry

Tumanov, S., Bulusu, V. and Kamphorst, J. J. (2015) Analysis of fatty acid metabolism using stable isotope tracers and mass spectrometry. In: Metallo, C. M. (ed.) Metabolic Analysis Using Stable Isotopes. Series: Methods in enzymology (561). Elsevier, pp. 197-217. ISBN 9780128022931 (doi: 10.1016/bs.mie.2015.05.017)

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Cells can synthesize fatty acids by ligating multiple acetyl units from acetyl-CoA. This is followed by desaturation and elongation reactions to produce a variety of fatty acids required for proper cellular functioning. Alternatively, exogenous lipid sources can contribute to cellular fatty acid pools. Here, we present a method based on incorporation of 13C-carbon from labeled substrates into fatty acids and subsequent mass spectrometry analysis. The resulting labeling patterns can be used to determine (1) 13C-enrichment of lipogenic acetyl-CoA, (2) the relative contributions of synthesis and uptake, and (3) absolute fatty acid fluxes. We begin by providing a background and general principles regarding the use of stable isotopes to study fatty acid metabolism. We then proceed with detailing procedures for sample preparation and both GC-MS and LC-MS analysis of isotope incorporation. Finally, we discuss the interpretation of the resulting fatty acid-labeling patterns.

Item Type:Book Sections
Glasgow Author(s) Enlighten ID:Tumanov, Dr Sergey and Kamphorst, Dr Jurre
Authors: Tumanov, S., Bulusu, V., and Kamphorst, J. J.
College/School:College of Medical Veterinary and Life Sciences > School of Cancer Sciences
Journal Name:Methods in Enzymology
ISSN (Online):1557-7988

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