| Autor: |
Varner, Erika L, Trefely, Sophie, Bartee, David, von Krusenstiern, Eliana, Izzo, Luke, Bekeova, Carmen, O’Connor, Roddy S, Seifert, Erin L., Wellen, Kathryn, Meier, Jordan L., Snyder, Nathaniel W. |
| Jazyk: |
angličtina |
| Rok vydání: |
2020 |
| DOI: |
10.1101/2020.03.15.992859 |
| Popis: |
Lysine lactoylation is a recently described protein post-translational modification (PTM). However, the biochemical pathways responsible for this acylation remain unclear. Two metabolite-dependent mechanisms have been proposed: enzymatic histone lysine lactoylation derived from lactoyl-coenzyme A (lactoyl-CoA), and non-enzymatic lysine lactoylation resulting from acyl-transfer via lactoyl-glutathione. While the former has precedent in the form of enzyme-catalyzed lysine acylation, the lactoyl-CoA metabolite has not been previously quantified in mammalian systems. Here we use liquid chromatography-high resolution mass spectrometry (LC-HRMS) together with a synthetic standard to detect and validate the presence of lactoyl-CoA in cell and tissue samples. Conducting a retrospective analysis of data from previously analyzed samples revealed the presence of lactoyl-CoA in diverse cell and tissue contexts. In addition, we describe a biosynthetic route to generate 13 C 3 15 N 1 -isotopically-labeled lactoyl-CoA, providing a co-eluting internal standard for analysis of this metabolite. These analyses enable estimation of lactoyl-CoA concentrations of 1.14×10 −8 pmol/cell in cell culture and 0.0172 pmol/mg tissue wet weight in mouse heart. These levels are similar to crotonyl-CoA, but between 20-350 times lower than major acyl-CoAs such as acetyl-, propionyl- and succinyl-CoA. Overall our studies provide the first quantitative measurements of lactoyl-CoA, and provide a methodological foundation for the interrogation of this novel metabolite in biology and disease. Highlights - Detection of lactoyl-CoA at picomole concentrations across tissues and cells - Lactoyl-CoA was detected at concentrations similar to crontonyl-CoA within HepG2 cells - Isotopically labeled 13 C 3 15 N 1 -lactoyl-CoA can be prepared by SILEC |
| Databáze: |
OpenAIRE |
| Externí odkaz: |
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