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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.


Madelaine C. Brearley

Zoe C.T.R. Daniel

Paul T. Loughna

Professor of Nutritional Biochemistry

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Professor of Nutritional Biochemstry


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.


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.

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
Print ISSN 2045-2322
Electronic ISSN 2045-2322
Publisher Nature Publishing Group
Peer Reviewed Peer Reviewed
Volume 10
Article Number 22177
Keywords Multidisciplinary
Public URL
Publisher URL
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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