Madelaine C. Brearley
The phosphoenolpyruvate carboxykinase (PEPCK) inhibitor, 3-mercaptopicolinic acid (3-MPA), induces myogenic differentiation in C2C12 cells
Brearley, Madelaine C.; Daniel, Zoe C.T.R.; Loughna, Paul T.; Parr, Tim; Brameld, John M.
Authors
Zoe C.T.R. Daniel
Paul T. Loughna
Professor TIM PARR TIM.PARR@NOTTINGHAM.AC.UK
PROFESSOR OF NUTRITIONAL BIOCHEMISTRY
Professor John Brameld JOHN.BRAMELD@NOTTINGHAM.AC.UK
PROFESSOR OF NUTRITIONAL BIOCHEMISTRY
Abstract
Phosphoenolpyruvate carboxykinase (PEPCK) is a gluconeogenic enzyme with a cytosolic (Pck1/PEPCK-C) and mitochondrial (Pck2/PEPCK-M) isoform. Here we investigate the effect of 3-mercaptopicolinic acid (3-MPA), a PEPCK inhibitor, on C2C12 muscle cells. We report that Pck2 mRNA is 50–5000-fold higher than Pck1 during C2C12 myogenesis, indicating Pck2 is the predominant PEPCK isoform. C2C12 cell proliferation was inhibited in a dose-dependent manner following 48h 3-MPA treatment (0.01–1mM). C2C12 myogenic differentiation was significantly induced following 3-MPA treatment (0.25, 0.5, 1mM) from day 0 of differentiation, demonstrated by increased creatine kinase activity, fusion index and myotube diameter; likewise, the myosin heavy chain (MyHC)-IIB isoform (encoded by Myh4) is an indicator of hypertrophy, and both porcine MYH4-promoter activity and endogenous Myh4 mRNA were also significantly induced. High doses (0.5 and/or 1mM) of 3-MPA reduced mRNA expression of Pck2 and genes associated with serine biosynthesis (Phosphoglycerate dehydrogenase, Phgdh; phosphoserine aminotransferase-1, Psat1) following treatment from days 0 and 4. To conclude, as Pck2/PEPCK-M is the predominant isoform in C2C12 cells, we postulate that 3-MPA promoted myogenic differentiation through the inhibition of PEPCK-M. However, we were unable to confirm that 3-MPA inhibited PEPCK-M enzyme activity as 3-MPA interfered with the PEPCK enzyme assay, particularly at 0.5 and 1mM.
Citation
Brearley, M. C., Daniel, Z. C., Loughna, P. T., Parr, T., & Brameld, J. M. (2020). The phosphoenolpyruvate carboxykinase (PEPCK) inhibitor, 3-mercaptopicolinic acid (3-MPA), induces myogenic differentiation in C2C12 cells. Scientific Reports, 10, Article 22177. https://doi.org/10.1038/s41598-020-79324-9
Journal Article Type | Article |
---|---|
Acceptance Date | Dec 2, 2020 |
Online Publication Date | Dec 17, 2020 |
Publication Date | Dec 17, 2020 |
Deposit Date | Dec 9, 2020 |
Publicly Available Date | Jan 5, 2021 |
Journal | Scientific Reports |
Electronic ISSN | 2045-2322 |
Publisher | Nature Publishing Group |
Peer Reviewed | Peer Reviewed |
Volume | 10 |
Article Number | 22177 |
DOI | https://doi.org/10.1038/s41598-020-79324-9 |
Keywords | Multidisciplinary |
Public URL | https://nottingham-repository.worktribe.com/output/5129246 |
Publisher URL | https://www.nature.com/articles/s41598-020-79324-9 |
Additional Information | Received: 11 February 2020; Accepted: 2 December 2020; First Online: 17 December 2020; : The authors declare no competing interests. |
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The phosphoenolpyruvate carboxykinase (PEPCK) inhibitor, 3-mercaptopicolinic acid (3-MPA), induces myogenic differentiation in C2C12 cells
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