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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Abstract
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 |
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Status: | Published |
Refereed: | Yes |
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 |
Publisher: | Elsevier |
ISSN: | 0076-6879 |
ISSN (Online): | 1557-7988 |
ISBN: | 9780128022931 |
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