Quantitation of fatty acyl-coenzyme As in mammalian cells by liquid chromatography-electrospray ionization tandem mass spectrometry*

Journal of Lipid Research

Alfred H. Merrill, Jr.¹  Kacee Sims²  Jeremy C. Allegood²  Elaine W. Wang³  Christopher A. Haynes³  M. Cameron Sullards³ 

[1] Chemistry and Biochemistry and the Parker H. Petit Institute for Bioengineering and Bioscience, Georgia Institute of Technology, Atlanta, GA 30332-0230;Chemistry and Biochemistry and the Parker H. Petit Institute for Bioengineering and Bioscience, Georgia Institute of Technology, Atlanta, GA 30332-0230;Schools of Biology, Georgia Institute of Technology, Atlanta, GA 30332-0230;

关键词: lipidomics;    metabolomics;    RAW264.7;    MCF7;    lignoceroyl-CoA;    nervonoyl-CoA;   

DOI  :  

来源: DOAJ

【 摘 要 】

Fatty acyl-CoAs participate in numerous cellular processes. This article describes a method for the quantitation of subpicomole amounts of long-chain and very-long-chain fatty acyl-CoAs by reverse-phase LC combined with electrospray ionization tandem mass spectrometry in positive ion mode with odd-chain-length fatty acyl-CoAs as internal standards. This method is applicable to a wide range of species [at least myristoyl- (C14:0-) to cerotoyl- (C26:0-) CoA] in modest numbers of cells in culture (∼106–107), with analyses of RAW264.7 cells and MCF7 cells given as examples. Analysis of these cells revealed large differences in fatty acyl-CoA amounts (12 ± 1.0 pmol/106 RAW264.7 cells vs. 80.4 ± 6.1 pmol/106 MCF7 cells) and subspecies distribution. Very-long-chain fatty acyl-CoAs with alkyl chain lengths > C20 constitute <10% of the total fatty acyl-CoAs of RAW264.7 cells versus >50% for MCF7 cells, which somewhat astonishingly contain approximately as much C24:0- and C26:0-CoAs as C16:0- and C18:0-CoAs and essentially equal amounts of C26:1- and C18:1-CoAs. This simple and robust method should facilitate the inclusion of this family of compounds in “lipidomics” and “metabolomics” studies.

【 授权许可】

Unknown