脂肪酸
化学
质谱法
碳同位素
生物化学
脂肪酸代谢
稳定同位素比值
同位素
脂肪酸合成
新陈代谢
色谱法
同位素标记
有机化学
物理
量子力学
总有机碳
作者
Sergey Tumanov,Vinay Bulusu,Jurre J. Kamphorst
出处
期刊:Methods in Enzymology
日期:2015-01-01
卷期号:: 197-217
被引量:36
标识
DOI:10.1016/bs.mie.2015.05.017
摘要
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.
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