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Serine is a natural ligand and allosteric activator of pyruvate kinase M2

Nature volume 491, pages 458–462 (2012)Cite this article

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A Corrigendum to this article was published on 17 April 2013

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Abstract

Cancer cells exhibit several unique metabolic phenotypes that are critical for cell growth and proliferation1. Specifically, they overexpress the M2 isoform of the tightly regulated enzyme pyruvate kinase (PKM2), which controls glycolytic flux, and are highly dependent on de novo biosynthesis of serine and glycine2. Here we describe a new rheostat-like mechanistic relationship between PKM2 activity and serine biosynthesis. We show that serine can bind to and activate human PKM2, and that PKM2 activity in cells is reduced in response to serine deprivation. This reduction in PKM2 activity shifts cells to a fuel-efficient mode in which more pyruvate is diverted to the mitochondria and more glucose-derived carbon is channelled into serine biosynthesis to support cell proliferation.

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Figure 1: Characterization of PKM-silenced HCT116 cells.
Figure 2: The effect of PKM silencing on glycolytic flux.
Figure 3: Serine and glycine deprivation changes glucose metabolism.
Figure 4: Serine is an allosteric activator of PKM2.

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Primary accessions

Protein Data Bank

Data deposits

Atomic coordinates and structure factors for the PKM2 crystal structures have been deposited in the Protein Data Bank (PDB) under accession code 4B2D.

Change history

  • 14 November 2012

    A url at the end of the Methods section was corrected.

  • 17 April 2013

    Nature 491, 458–462 (2012); doi:10.1038/nature11540 In Fig. 1a of this Letter, the western blot originally published in the PKM2 panel was a re-probe of the blot used in the PKM1 panel and the signal picked up was from the first antibody (PKM1) and not from PKM2. We have repeated the experiment on three different membranes with the antibodies used in the original paper for PKM1, PKM2 and total PKM, using actin as the loading control (Fig.

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Acknowledgements

The work performed at the Beatson Institute for Cancer Research was supported by Cancer Research UK. We thank D. Sumpton for technical support with two-dimensional gel electrophoresis and N. Thompson, N. Wallis and M. Jones for comments provided during manuscript preparation. We would also like to thank D. M. Sabatini for the Scramble shRNA plasmid used as a control (shCntrla) and the Structural Genomics Consortium for providing us with the PKM2 expression plasmid from their collection. We thank A. King for editorial work and S. Tardito for graphical help.

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Author notes

  1. Christian Frezza

    Present address: Present address: MRC Cancer Cell Unit, Hutchison/MRC Research Centre, Hills Road, Cambridge, CB2 0XZ, UK.,

  2. Barbara Chaneton and Petra Hillmann: These authors contributed equally to this work.

Authors and Affiliations

  1. Cancer Research UK, The Beatson Institute for Cancer Research, Switchback Road, Glasgow G61 1BD, Scotland, UK,

    Barbara Chaneton, Liang Zheng, Oliver D. K. Maddocks, Karen H. Vousden, Christian Frezza & Eyal Gottlieb

  2. Astex Pharmaceuticals, 436 Cambridge Science Park, Milton Road, Cambridge CB4 0QA, UK,

    Petra Hillmann, Agnès C. L. Martin, Joseph E. Coyle, Finn P. Holding & Marc O’Reilly

  3. Institute of Molecular, Cell and Systems Biology, College of Medical, Veterinary and Life Sciences, Joseph Black Building, B3.09, University of Glasgow, Glasgow G12 8QQ, Scotland, UK,

    Achuthanunni Chokkathukalam & Andris Jankevics

  4. Groningen Bioinformatics Centre, Groningen Biomolecular Sciences and Biotechnology Institute, University of Groningen, Groningen, 9747 AG, The Netherlands

    Andris Jankevics

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Contributions

M.O. and E.G. conceived the project and wrote the manuscript with the help of B.C., P.H. and C.F. L.Z., B.C. and C.F. performed the LC–MS assay and analysed the raw data. A.C. and A.J. analysed the LC–MS data and identified the different isotopomers of each metabolite. A.C.L.M. performed the in vitro enzymatic activity, J.E.C. performed the ITC, M.O. generated the point mutant constructs, purified the proteins and solved the crystal structure. F.P.H. performed the LC–MS validation of the point mutant constructs. O.D.K.M. and K.H.V. performed, analysed and discussed the long-term serine and glycine starvation experiment. B.C. and P.H. generated and characterized the cell lines and performed all other experiments and data analysis. All the authors discussed the results and commented on the manuscript.

Corresponding authors

Correspondence to Marc O’Reilly or Eyal Gottlieb.

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Competing interests

E.G. is a consultant of Astex Pharmaceuticals.

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Chaneton, B., Hillmann, P., Zheng, L. et al. Serine is a natural ligand and allosteric activator of pyruvate kinase M2. Nature 491, 458–462 (2012). https://doi.org/10.1038/nature11540

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