Cancer-associated mutations in Human Pyruvate Kinase M2 Impair Enzyme Activity

Overview

Journal FEBS Lett

Specialty Biochemistry

Date 2019 Oct 24

PMID 31642061

Citations 13

Authors

Vivian M Liu

Andrea J Howell

Aaron M Hosios

Zhaoqi Li

William J Israelsen

Matthew G Vander Heiden

Affiliations

Soon will be listed here.

Abstract

Mammalian pyruvate kinase catalyzes the final step of glycolysis, and its M2 isoform (PKM2) is widely expressed in proliferative tissues. Mutations in PKM2 are found in some human cancers; however, the effects of these mutations on enzyme activity and regulation are unknown. Here, we characterized five cancer-associated PKM2 mutations, occurring at various locations on the enzyme, with respect to substrate kinetics and activation by the allosteric activator fructose-1,6-bisphosphate (FBP). The mutants exhibit reduced maximal velocity, reduced substrate affinity, and/or altered activation by FBP. The kinetic parameters of five additional PKM2 mutants that have been used to study enzyme function or regulation also demonstrate the deleterious effects of mutations on PKM2 function. Our findings indicate that PKM2 is sensitive to many amino acid changes and support the hypothesis that decreased PKM2 activity is selected for in rapidly proliferating cells.

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References

Bollenbach T, Mesecar A, Nowak T . Role of lysine 240 in the mechanism of yeast pyruvate kinase catalysis. Biochemistry. 1999; 38(28):9137-45. DOI: 10.1021/bi990690n. View

Blair J, Walker R . Rat liver pyruvate kinase: influence of ligands on activity and fructose 1,6-bisphosphate binding. Arch Biochem Biophys. 1984; 232(1):202-13. DOI: 10.1016/0003-9861(84)90536-8. View

Yang W, Xia Y, Ji H, Zheng Y, Liang J, Huang W . Nuclear PKM2 regulates β-catenin transactivation upon EGFR activation. Nature. 2011; 480(7375):118-22. PMC: 3235705. DOI: 10.1038/nature10598. View

Koler R, VANBELLINGHEN P . The mechanism of precursor modulation of human pyruvate kinase I by fructose diphosphate. Adv Enzyme Regul. 1968; 6:127-42. DOI: 10.1016/0065-2571(68)90010-1. View

Allali-Hassani A, Wasney G, Chau I, Hong B, Senisterra G, Loppnau P . A survey of proteins encoded by non-synonymous single nucleotide polymorphisms reveals a significant fraction with altered stability and activity. Biochem J. 2009; 424(1):15-26. DOI: 10.1042/BJ20090723. View

Jiang Y, Wang Y, Wang T, Hawke D, Zheng Y, Li X . PKM2 phosphorylates MLC2 and regulates cytokinesis of tumour cells. Nat Commun. 2014; 5:5566. PMC: 4259466. DOI: 10.1038/ncomms6566. View

MUIRHEAD H, Clayden D, Barford D, Lorimer C, Fothergill-Gilmore L, Schiltz E . The structure of cat muscle pyruvate kinase. EMBO J. 1986; 5(3):475-81. PMC: 1166788. DOI: 10.1002/j.1460-2075.1986.tb04236.x. View

Cortes-Cros M, Hemmerlin C, Ferretti S, Zhang J, Gounarides J, Yin H . M2 isoform of pyruvate kinase is dispensable for tumor maintenance and growth. Proc Natl Acad Sci U S A. 2012; 110(2):489-94. PMC: 3545759. DOI: 10.1073/pnas.1212780110. View

Hitosugi T, Kang S, Vander Heiden M, Chung T, Elf S, Lythgoe K . Tyrosine phosphorylation inhibits PKM2 to promote the Warburg effect and tumor growth. Sci Signal. 2009; 2(97):ra73. PMC: 2812789. DOI: 10.1126/scisignal.2000431. View

10.

Dayton T, Gocheva V, Miller K, Bhutkar A, Lewis C, Bronson R . Isoform-specific deletion of PKM2 constrains tumor initiation in a mouse model of soft tissue sarcoma. Cancer Metab. 2018; 6:6. PMC: 5977456. DOI: 10.1186/s40170-018-0179-2. View

11.

Lau A, Israelsen W, Roper J, Sinnamon M, Georgeon L, Dayton T . PKM2 is not required for colon cancer initiated by APC loss. Cancer Metab. 2017; 5:10. PMC: 5707917. DOI: 10.1186/s40170-017-0172-1. View

12.

Macpherson J, Theisen A, Masino L, Fets L, Driscoll P, Encheva V . Functional cross-talk between allosteric effects of activating and inhibiting ligands underlies PKM2 regulation. Elife. 2019; 8. PMC: 6636998. DOI: 10.7554/eLife.45068. View

13.

Morgan H, OReilly F, Wear M, ONeill J, Fothergill-Gilmore L, Hupp T . M2 pyruvate kinase provides a mechanism for nutrient sensing and regulation of cell proliferation. Proc Natl Acad Sci U S A. 2013; 110(15):5881-6. PMC: 3625322. DOI: 10.1073/pnas.1217157110. View

14.

Cardenas J, Blachly E, Ceccotti P, Dyson R . Properties of chicken skeletal muscle pyruvate kinase and a proposal for its evolutionary relationship to the other avian and mammalian isozymes. Biochemistry. 1975; 14(10):2247-52. DOI: 10.1021/bi00681a032. View

15.

Wang Y, Israelsen W, Lee D, Yu V, Jeanson N, Clish C . Cell-state-specific metabolic dependency in hematopoiesis and leukemogenesis. Cell. 2014; 158(6):1309-1323. PMC: 4197056. DOI: 10.1016/j.cell.2014.07.048. View

16.

Clower C, Chatterjee D, Wang Z, Cantley L, Vander Heiden M, Krainer A . The alternative splicing repressors hnRNP A1/A2 and PTB influence pyruvate kinase isoform expression and cell metabolism. Proc Natl Acad Sci U S A. 2010; 107(5):1894-9. PMC: 2838216. DOI: 10.1073/pnas.0914845107. View

17.

Lu Z, Hunter T . Metabolic Kinases Moonlighting as Protein Kinases. Trends Biochem Sci. 2018; 43(4):301-310. PMC: 5879014. DOI: 10.1016/j.tibs.2018.01.006. View

18.

Taylor C, Bailey E . Activation of liver pyruvate kinase by fructose 1,6-diphosphate. Biochem J. 1967; 102(2):32C-33C. PMC: 1270301. DOI: 10.1042/bj1020032c. View

19.

Christofk H, Vander Heiden M, Harris M, Ramanathan A, Gerszten R, Wei R . The M2 splice isoform of pyruvate kinase is important for cancer metabolism and tumour growth. Nature. 2008; 452(7184):230-3. DOI: 10.1038/nature06734. View

20.

Yang P, Li Z, Fu R, Wu H, Li Z . Pyruvate kinase M2 facilitates colon cancer cell migration via the modulation of STAT3 signalling. Cell Signal. 2014; 26(9):1853-62. DOI: 10.1016/j.cellsig.2014.03.020. View