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Pyruvate kinase M2 is a phosphotyrosine-binding protein

Nature volume 452, pages 181–186 (2008)Cite this article

Abstract

Growth factors stimulate cells to take up excess nutrients and to use them for anabolic processes. The biochemical mechanism by which this is accomplished is not fully understood but it is initiated by phosphorylation of signalling proteins on tyrosine residues. Using a novel proteomic screen for phosphotyrosine-binding proteins, we have made the observation that an enzyme involved in glycolysis, the human M2 (fetal) isoform of pyruvate kinase (PKM2), binds directly and selectively to tyrosine-phosphorylated peptides. We show that binding of phosphotyrosine peptides to PKM2 results in release of the allosteric activator fructose-1,6-bisphosphate, leading to inhibition of PKM2 enzymatic activity. We also provide evidence that this regulation of PKM2 by phosphotyrosine signalling diverts glucose metabolites from energy production to anabolic processes when cells are stimulated by certain growth factors. Collectively, our results indicate that expression of this phosphotyrosine-binding form of pyruvate kinase is critical for rapid growth in cancer cells.

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Figure 1: Proteomic screen identifies pyruvate kinase as a novel phosphotyrosine-binding protein.
Figure 2: Phosphopeptide binding is specific to the M2 isoform of pyruvate kinase and involves lysine 433 near the FBP binding pocket.
Figure 3: Phosphotyrosine peptide catalyses the release of FBP from PKM2.
Figure 4: Phosphopeptide binding results in inhibition of PKM2 activity.
Figure 5: The phosphopeptide binding ability of PKM2 is important for cell proliferation.

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Protein Data Bank

Data deposits

PKM2 structure files are deposited in the Protein Data Bank under accession codes 3BJT (apo form) and 3BJF (FBP-bound form).

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Acknowledgements

We thank W. Hahn for providing the lentiviral shRNA. We thank M. Liu for technical assistance. We thank J. Lee for help in developing the affinity purification techniques. M.G.V.H. is a Damon Runyon Fellow supported by the Damon Runyon Cancer Research Foundation. This research was supported by funding to L.C.C. from the National Institutes of Health.

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Authors and Affiliations

  1. Department of Systems Biology,,

    Heather R. Christofk, Matthew G. Vander Heiden, Ning Wu & Lewis C. Cantley

  2. Department of Pathology, Harvard Medical School, Boston, Massachusetts 02115, USA,

    John M. Asara

  3. Dana Farber Cancer Institute, Boston, Massachusetts 02115, USA,

    Matthew G. Vander Heiden

  4. Division of Signal Transduction, Beth Israel Deaconess Medical Center, Boston, Massachusetts 02115, USA,

    John M. Asara & Lewis C. Cantley

Authors
  1. Heather R. Christofk
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  2. Matthew G. Vander Heiden
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  3. Ning Wu
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  4. John M. Asara
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  5. Lewis C. Cantley
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Corresponding author

Correspondence to Lewis C. Cantley.

Supplementary information

Supplementary Information

The file contains Supplementary Discussion; Supplementary Figures 1-4 with Legends illustrating data from control experiments; and Supplementary Tables 1- 2 showing the x-ray crystallography refinement statistics and a list of the phosphopeptides used to assess PKM2 activity. (PDF 282 kb)

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Christofk, H., Vander Heiden, M., Wu, N. et al. Pyruvate kinase M2 is a phosphotyrosine-binding protein. Nature 452, 181–186 (2008). https://doi.org/10.1038/nature06667

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