• A Corrigendum to this article was published on 17 April 2013

This article has been updated

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.

Access optionsAccess options

Rent or Buy article

Get time limited or full article access on ReadCube.

from$8.99

All prices are NET prices.

Change history

  • 14 November 2012

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

Accessions

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.

References

  1. 1.

    , & Targeting metabolic transformation for cancer therapy. Nature Rev. Cancer 10, 267–277 (2010)

  2. 2.

    & Rocking cell metabolism: revised functions of the key glycolytic regulator PKM2 in cancer. Trends Biochem. Sci. 37, 309–316 (2012)

  3. 3.

    et al. The M2 splice isoform of pyruvate kinase is important for cancer metabolism and tumour growth. Nature 452, 230–233 (2008)

  4. 4.

    & Genes of glycolysis are ubiquitously overexpressed in 24 cancer classes. Genomics 84, 1014–1020 (2004)

  5. 5.

    , , & Pyruvate kinase type M2 and its role in tumor growth and spreading. Semin. Cancer Biol. 15, 300–308 (2005)

  6. 6.

    & Nutrient and hormonal regulation of pyruvate kinase gene expression. Biochem. J. 337, 1–11 (1999)

  7. 7.

    et al. Functional genomics reveal that the serine synthesis pathway is essential in breast cancer. Nature 476, 346–350 (2011)

  8. 8.

    et al. Phosphoglycerate dehydrogenase diverts glycolytic flux and contributes to oncogenesis. Nature Genet. 43, 869–874 (2011)

  9. 9.

    et al. Enhanced serine production by bone metastatic breast cancer cells stimulates osteoclastogenesis. Breast Cancer Res. Treat. 125, 421–430 (2011)

  10. 10.

    et al. Evidence for an alternative glycolytic pathway in rapidly proliferating cells. Science 329, 1492–1499 (2010)

  11. 11.

    , & Structural basis for tumor pyruvate kinase M2 allosteric regulation and catalysis. Biochemistry 44, 9417–9429 (2005)

  12. 12.

    et al. Tyrosine phosphorylation inhibits PKM2 to promote the Warburg effect and tumor growth. Sci. Signal. 2, ra73 (2009)

  13. 13.

    , , , & Pyruvate kinase M2 is a phosphotyrosine-binding protein. Nature 452, 181–186 (2008)

  14. 14.

    , , & In vivo regulation of monomer-tetramer conversion of pyruvate kinase subtype M2 by glucose is mediated via fructose 1,6-bisphosphate. J. Biol. Chem. 266, 16842–16846 (1991)

  15. 15.

    & Similarities between pyruvate kinase from human placenta and tumours. FEBS Lett. 37, 281–284 (1973)

  16. 16.

    , , , & Structural and kinetic differences between the M2 type pyruvate kinases from lung and various tumors. Biomed. Biochim. Acta 42, S278–S282 (1983)

  17. 17.

    et al. Pyruvate kinase M2 promotes de novo serine synthesis to sustain mTORC1 activity and cell proliferation. Proc. Natl Acad. Sci. USA 109, 6904–6909 (2012)

  18. 18.

    & Fragment screening using X-ray crystallography. Top. Curr. Chem. 317, 33–59 (2012)

  19. 19.

    et al. A survey of proteins encoded by non-synonymous single nucleotide polymorphisms reveals a significant fraction with altered stability and activity. Biochem. J. 424, 15–26 (2009)

  20. 20.

    , & Interchange of amino acids between tumor and host. Biochem. Med. Metab. Biol. 48, 1–7 (1992)

  21. 21.

    , , , & Nitrogen metabolism in tumor bearing mice. Arch. Biochem. Biophys. 268, 667–675 (1989)

  22. 22.

    , & The allosteric regulation of pyruvate kinase. FEBS Lett. 389, 15–19 (1996)

  23. 23.

    et al. Inhibition of pyruvate kinase M2 by reactive oxygen species contributes to cellular antioxidant responses. Science 334, 1278–1283 (2011)

  24. 24.

    et al. l-serine in disease and development. Biochem. J. 371, 653–661 (2003)

  25. 25.

    , , , & Simultaneous determination of multiple intracellular metabolites in glycolysis, pentose phosphate pathway and tricarboxylic acid cycle by liquid chromatography-mass spectrometry. J. Chromatogr. A 1147, 153–164 (2007)

  26. 26.

    & Therapeutic compositions and related methods of use. PTC patent application WO/2010/118063. (2010)

  27. 27.

    et al. Evaluation of substituted N,N′-diarylsulfonamides as activators of the tumor cell specific M2 isoform of pyruvate kinase. J. Med. Chem. 53, 1048–1055 (2010)

  28. 28.

    . The CCP4 suite: programs for protein crystallography. Acta Crystallogr. D 50, 760–763 (1994)

  29. 29.

    et al. Automated protein-ligand crystallography for structure-based drug design. Chem Med Chem 1, 827–838 (2006)

  30. 30.

    Free R value: a novel statistical quantity for assessing the accuracy of crystal structures. Nature 355, 472–475 (1992)

  31. 31.

    & Improved R factors for diffraction data analysis in macromolecular crystallography. Nature Struct. Biol. 4, 269–275 (1997)

  32. 32.

    & On the use of the merging R factor as a quality indicator for X-ray data. J. Appl. Crystallogr. 30, 203–205 (1997)

  33. 33.

    Global indicators of X-ray data quality. J. Appl. Crystallogr. 34, 130–135 (2001)

  34. 34.

    et al. A common open representation of mass spectrometry data and its application to proteomics research. Nature Biotechnol. 22, 1459–1466 (2004)

  35. 35.

    , , , & ProteoWizard: open source software for rapid proteomics tools development. Bioinformatics 24, 2534–2536 (2008)

  36. 36.

    , & Highly sensitive feature detection for high resolution LC/MS. BMC Bioinformatics 9, 504 (2008)

  37. 37.

    , , , & XCMS: processing mass spectrometry data for metabolite profiling using nonlinear peak alignment, matching, and identification. Anal. Chem. 78, 779–787 (2006)

  38. 38.

    , , , & Correction of 13C mass isotopomer distributions for natural stable isotope abundance. J. Mass Spectrom. 31, 255–262 (1996)

  39. 39.

    , , , & PeakML/mzMatch: a file format, Java library, R library, and tool-chain for mass spectrometry data analysis. Anal. Chem. 83, 2786–2793 (2011)

Download references

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.

Author information

Author notes

    • Barbara Chaneton
    •  & Petra Hillmann

    These authors contributed equally to this work.

    • Christian Frezza

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

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

Authors

  1. Search for Barbara Chaneton in:

  2. Search for Petra Hillmann in:

  3. Search for Liang Zheng in:

  4. Search for Agnès C. L. Martin in:

  5. Search for Oliver D. K. Maddocks in:

  6. Search for Achuthanunni Chokkathukalam in:

  7. Search for Joseph E. Coyle in:

  8. Search for Andris Jankevics in:

  9. Search for Finn P. Holding in:

  10. Search for Karen H. Vousden in:

  11. Search for Christian Frezza in:

  12. Search for Marc O’Reilly in:

  13. Search for Eyal Gottlieb in:

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.

Competing interests

E.G. is a consultant of Astex Pharmaceuticals.

Corresponding authors

Correspondence to Marc O’Reilly or Eyal Gottlieb.

Supplementary information

PDF files

  1. 1.

    Supplementary Information

    This file contains a Supplementary Discussion, Supplementary Figures 1-9 and Supplementary Tables 1-2.

About this article

Publication history

Received

Accepted

Published

DOI

https://doi.org/10.1038/nature11540

Further reading

Comments

By submitting a comment you agree to abide by our Terms and Community Guidelines. If you find something abusive or that does not comply with our terms or guidelines please flag it as inappropriate.

Newsletter Get the most important science stories of the day, free in your inbox. Sign up for Nature Briefing