siRNA knockdown, neuroblastoma cells, N-acylamide, arachidonic acid, brain lipids, lipids, eicosanoids, mass spectrometry, palmitoylation, oleamide
Digital Object Identifier (DOI)
Long-chain fatty acid amides are signaling lipids found in mammals and other organisms; however, details of the metabolic pathways for the N-acylglycines and primary fatty acid amides (PFAMs) have remained elusive. Heavy-labeled precursor and subtraction lipidomic experiments in mouse neuroblastoma N18TG2 cells, a model cell line for the study of fatty acid amide metabolism, establish the biosynthetic pathways for the N-acylglycines and the PFAMs. We provide evidence that the N-acylglycines are formed by a long-chain specific glycine-conjugating enzyme, glycine N-acyltransferase-like 3 (GLYATL3). siRNA knockdown of GLYATL3 in the N18TG2 cells resulted in a decrease in the levels of the N-acylglycines and the PFAMs. This is the first report of an enzyme responsible for long-chain N-acylglycine production in cellula. The production of the PFAMs in N18TG2 cells was reported to occur by the oxidative cleavage of the N-acylglycines, as catalyzed by peptidylglycine α-amidating monooxygenase (PAM). siRNA knockdown of PAM resulted in an accumulation of [(13)C18]N-oleoylglycine and decreased levels of [(13)C18]oleamide when the N18TG2 cells were grown in the presence of [(13)C18]oleic acid. The addition of [1-(13)C]palmitate to the N18TG2 cell growth media led to the production of a family of [1-(13)C]palmitoylated fatty acid amides, consistent with the biosynthetic pathways detailed herein.
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Citation / Publisher Attribution
Journal of Lipid Research, v. 57, p. 781-790
This research was originally published in the Journal of Lipid Research. Jeffries, K. A., Dempsey, D. R., Farrell, E. K., Anderson, R. L., Garbade, G. J., Gurina, T. S., Gruhonjic, I., Gunderson, C. A., and Merkler, D. J. Glycine N-acyltransferase-like 3 is Responsible for Long-chain N-acylglycine Formation in N(18)TG(2) Cells. J. Lipid Res. 2016; Vol: 781-790. © the American Society for Biochemistry and Molecular.
Scholar Commons Citation
Jeffries, Kristen A; Dempsey, Daniel R; Farrell, Emma K; Anderson, Ryan L; Garbade, Gabrielle J; Gurina, Tatyana S; Gruhonjic, Imran; Gunderson, Carly A; and Merkler, David J, "Glycine N-acyltransferase-like 3 is Responsible for Long-chain N-acylglycine Formation in N18TG2 Cells" (2016). Chemistry Faculty Publications. 23.