2-Hydroxyglutarate (2-HG) is a hypoxic metabolite with potentially important epigenetic signaling roles. The mechanisms underlying 2-HG generation are poorly understood, but evidence suggests a potential regulatory role for the sirtuin family of lysine deacetylases. Thus, we hypothesized that the acetylation status of the major 2-HG-generating enzymes [lactate dehydrogenase (LDH), isocitrate dehydrogenase (IDH) and malate dehydrogenase (MDH)] may govern their 2-HG-generating activity. In vitro acetylation of these enzymes, with confirmation by western blotting, mass spectrometry, reversibility by recombinant sirtuins and an assay for global lysine occupancy, yielded no effect on 2-HG-generating activity. In addition, while elevated 2-HG in hypoxia is associated with the activation of lysine deacetylases, we found that mice lacking mitochondrial SIRT3 exhibited hyperacetylation and elevated 2-HG. These data suggest that there is no direct link between enzyme acetylation and 2-HG production. Furthermore, our observed effects of in vitro acetylation on the canonical activities of IDH, MDH and LDH appeared to contrast with previous findings wherein acetyl-mimetic lysine mutations resulted in the inhibition of these enzymes. Overall, these data suggest that a causal relationship should not be assumed between acetylation of metabolic enzymes and their activities, canonical or otherwise.
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August 2017
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The distribution of haemagglutinin (green) and β-catenin (red) in MDCK cells overexpressing Akt1-HA. In this issue of the Biochemical Journal, Castañeda et al. report on the suppression of Akt/β-catenin-mediated cell proliferation by the the inhibition of 14-3-3ζ expression (see pages 2679–2689).
Research Article|
August 10 2017
Potential mechanisms linking SIRT activity and hypoxic 2-hydroxyglutarate generation: no role for direct enzyme (de)acetylation
Sergiy M. Nadtochiy;
Sergiy M. Nadtochiy
1Department of Anesthesiology, University of Rochester Medical Center, Rochester, NY 14642, U.S.A.
2Department of Neuroscience, University of Rochester Medical Center, Rochester, NY, U.S.A.
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Yves T. Wang;
Yves T. Wang
1Department of Anesthesiology, University of Rochester Medical Center, Rochester, NY 14642, U.S.A.
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Jimmy Zhang;
Jimmy Zhang
3Department of Pharmacology and Physiology, University of Rochester Medical Center, Rochester, NY, U.S.A.
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Keith Nehrke;
Keith Nehrke
4Department of Medicine, University of Rochester Medical Center, Rochester, NY, U.S.A.
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Xenia Schafer;
Xenia Schafer
5Department of Biochemistry, University of Rochester Medical Center, Rochester, NY, U.S.A.
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Kevin Welle;
Kevin Welle
6Department of Proteomics Core Facility, University of Rochester Medical Center, Rochester, NY, U.S.A.
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Sina Ghaemmaghami;
Sina Ghaemmaghami
7Department of Biology, University of Rochester Medical Center, Rochester, NY, U.S.A.
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Josh Munger;
Josh Munger
5Department of Biochemistry, University of Rochester Medical Center, Rochester, NY, U.S.A.
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Paul S. Brookes
1Department of Anesthesiology, University of Rochester Medical Center, Rochester, NY 14642, U.S.A.
Correspondence: Paul S. Brookes (paul_brookes@urmc.rochester.edu)
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Publisher: Portland Press Ltd
Received:
May 19 2017
Revision Received:
June 28 2017
Accepted:
July 03 2017
Accepted Manuscript online:
July 03 2017
Online ISSN: 1470-8728
Print ISSN: 0264-6021
© 2017 The Author(s). Published by Portland Press Limited on behalf of the Biochemical Society
2017
Biochem J (2017) 474 (16): 2829–2839.
Article history
Received:
May 19 2017
Revision Received:
June 28 2017
Accepted:
July 03 2017
Accepted Manuscript online:
July 03 2017
Citation
Sergiy M. Nadtochiy, Yves T. Wang, Jimmy Zhang, Keith Nehrke, Xenia Schafer, Kevin Welle, Sina Ghaemmaghami, Josh Munger, Paul S. Brookes; Potential mechanisms linking SIRT activity and hypoxic 2-hydroxyglutarate generation: no role for direct enzyme (de)acetylation. Biochem J 15 August 2017; 474 (16): 2829–2839. doi: https://doi.org/10.1042/BCJ20170389
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