title: Analysis of Fatty Acid Metabolism Using Stable Isotope Tracers and Mass Spectrometry.
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 (13)C-carbon from labeled substrates into fatty acids and subsequent mass spectrometry analysis. The resulting labeling patterns can be used to determine (1) (13)C-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.
authors.affiliation: Cancer Research UK Beatson Institute & Institute of Cancer Sciences, University of Glasgow, Glasgow, United Kingdom.
authors.forename: Sergey
authors.initials: S
authors.lastname: Tumanov
authors.affiliation: Cancer Research UK Beatson Institute & Institute of Cancer Sciences, University of Glasgow, Glasgow, United Kingdom.
authors.forename: Vinay
authors.initials: V
authors.lastname: Bulusu
authors.affiliation: Cancer Research UK Beatson Institute & Institute of Cancer Sciences, University of Glasgow, Glasgow, United Kingdom. Electronic address: jurre.kamphorst@glasgow.ac.uk.
authors.forename: Jurre J
authors.initials: JJ
authors.lastname: Kamphorst
mesh_terms: D000105:Acetyl Coenzyme A / Q000378:metabolism; D000818:Animals; D002247:Carbon Isotopes; D005227:Fatty Acids / Q000378:metabolism*; D006801:Humans; D007553:Isotope Labeling / Q000379:methods*; D050356:Lipid Metabolism; D013058:Mass Spectrometry / Q000379:methods*
chemical_list: D002247:Carbon Isotopes; D005227:Fatty Acids; D000105:Acetyl Coenzyme A
keywords: Acetyl-CoA; De novo lipogenesis; Fatty acid metabolism; Mass spectrometry; Stable isotopes; Tracing