title: Development of a Method for the Determination of Acyl-CoA Compounds by Liquid Chromatography Mass Spectrometry to Probe the Metabolism of Fatty Acids.
abstract: Acyl-Coenzyme As (acyl-CoAs) are a group of activated fatty acid molecules participating in multiple cellular processes including lipid synthesis, oxidative metabolism of fatty acids to produce ATP, transcriptional regulation, and protein post-translational modification. Quantification of cellular acyl-CoAs is challenging due to their instability in aqueous solutions and lack of blank matrices. Here we demonstrate an LC-MS/MS analytical method which allows for absolute quantitation with broad coverage of cellular acyl-CoAs. This assay was applied to profile endogenous acyl-CoAs under the challenge of a variety of dietary fatty acids in prostate and hepatic cells. Additionally, this approach allowed for detection of multiple fatty acid metabolic processes including the biogenesis of acyl-CoAs, and their elongation, degradation, and desaturation. Hierarchical clustering in the remodeling of acyl-CoA profiles revealed a fatty-acid-specific pattern across all tested cell lines, which provides a valuable reference for making predictions in other cell models. Individual acyl-CoAs were identified which were altered differentially by exogenous fatty acids in divergent tumorigenicity states of cells. These findings demonstrate the power of acyl-CoA profiling toward understanding the mechanisms for the progression of tumors or other diseases in response to fatty acids.
authors.affiliation: Department of Pharmaceutical and Biomedical Sciences, University of Georgia , 250 West Green Street, Athens, Georgia 30602, United States.
authors.forename: Xiangkun
authors.initials: X
authors.lastname: Yang
authors.affiliation: Department of Pharmaceutical and Biomedical Sciences, University of Georgia , 250 West Green Street, Athens, Georgia 30602, United States.
authors.forename: Yongjie
authors.initials: Y
authors.lastname: Ma
authors.affiliation: Department of Analytical Chemistry, School of Pharmacy, Shenyang Pharmaceutical University , 103 Wenhua Road, Shenyang, 110016, China.
authors.forename: Ning
authors.initials: N
authors.lastname: Li
authors.affiliation: Department of Pharmaceutical and Biomedical Sciences, University of Georgia , 250 West Green Street, Athens, Georgia 30602, United States.
authors.forename: Houjian
authors.initials: H
authors.lastname: Cai
authors.affiliation: Department of Pharmaceutical and Biomedical Sciences, University of Georgia , 250 West Green Street, Athens, Georgia 30602, United States.
authors.forename: Michael G
authors.identifier: 0000-0003-0626-3234
authors.initials: MG
authors.lastname: Bartlett
mesh_terms: D000214:Acyl Coenzyme A / Q000737:chemistry* / Q000378:metabolism*; D002851:Chromatography, High Pressure Liquid; D005227:Fatty Acids / Q000032:analysis* / Q000378:metabolism*; D006801:Humans; D013058:Mass Spectrometry; D015394:Molecular Structure; D014407:Tumor Cells, Cultured
chemical_list: D000214:Acyl Coenzyme A; D005227:Fatty Acids