Professor Grahame Hardie FRS, FRSE, FMedSci
The AMP-activated protein kinase (AMPK) cascade was first defined in our laboratory. Almost all energy-consuming reactions in the cell are powered by the high ratio of ATP to ADP, and when energy stress causes this ratio to fall, the AMPK system is switched on. Due to the adenylate kinase reaction, any increase in the ADP:ATP ratio is accompanied by a large rise in AMP, and this is the primary signal that triggers AMPK activation.
AMPK occurs in essentially all eukaryotes as heterotrimers with catalytic alpha and regulatory beta and gamma subunits (Figure 1).
AMP and/or ADP binds to one or more of three sites formed by the CBS repeats on the gamma subunit, causing conformational changes that have three effects, all antagonized by binding of ATP:
- promoting phosphorylation of the alpha subunit at the activating site, Thr172, by the upstream kinase LKB1;
- inhibiting dephosphorylation at Thr172 by protein phosphatases – this effect can be mimicked by ADP;
- allosteric activation of the kinase already phosphorylated on Thr172.
This complex mechanism allows AMPK to act as an ultrasensitive sensor that monitors cellular AMP:ATP and ADP:ATP ratios (Figure 2). AMPK can also be activated by a rise in cellular calcium ions, which switch on the alternative upstream kinase, the calcium/calmodulin-dependent kinase kinase, CaMKK-beta.
Once activated by falling energy status or rising calcium ions, AMPK switches on ATP-producing catabolic pathways, while switching off ATP-consuming processes, including cell growth and proliferation. The AMPK system has also been found to play a key role in modulating energy balance at the whole body level, by mediating effects of hormones such as leptin, adiponectin and ghrelin that regulate food intake and energy expenditure. It is responsible for many of the acute metabolic changes and the longer-term metabolic adaptations of muscle to regular exercise, and may explain the protective effects of regular exercise on the development of obesity and type 2 diabetes. By reducing fat storage in cells, AMPK may also mediate the insulin-sensitizing effects of metformin, an anti-diabetic drug currently prescribed to over 100 million people worldwide.
Our finding in 2003 that LKB1 was the upstream kinase that phosphorylates Thr172 introduced a link between AMPK and cancer. We are currently investigating the role of AMPK in cancer, and whether AMPK activation can explain the apparent protective effects of metformin in development of the disease.
Recently, we discovered that salicylate, a natural product that is the major breakdown product of aspirin, activates AMPK by direct binding to the carbohydrate-binding module on the beta subunit. It is becoming clear that AMPK modulates metabolism in cells of the immune system, and we are interested in the possibility that AMPK may mediate some of the anti-inflammatory effects of aspirin and other salicylate-based drugs.
Zong, Y, Zhang, C-S, Li, M, Wang, W, Wang, Z, Hawley, SA, Ma, T, Feng, J-W, Tian, X, Qi, Q, Wu, Y-Q, Zhang, C, Ye, Z, Lin, S-Y, Piao, H-L, Hardie, DG & Lin, S-C 2019, 'Hierarchical activation of compartmentalized pools of AMPK depends on severity of nutrient or energy stress' Cell Research. https://doi.org/10.1038/s41422-019-0163-6
Vara Ciruelos, D, Dandapani, M, Russell, F, Grzes, KM, Atrih, A, Foretz, M, Viollet, B, Lamont, D, Cantrell, D & Hardie, G 2019, 'Phenformin, but not metformin, delays development of T-cell acute lymphoblastic leukemia/lymphoma via cell-autonomous AMPK activation' Cell Reports, vol. 27, no. 3, pp. 690-698.e4. https://doi.org/10.1016/j.celrep.2019.03.067
Lin, S-C & Hardie, D 2018, 'AMPK: Sensing Glucose as well as Cellular Energy Status' Cell Metabolism, vol. 27, no. 2, pp. 299-313. https://doi.org/10.1016/j.cmet.2017.10.009
Fyffe, FA, Hawley, SA, Gray, A & Hardie, DG 2018, Cell-Free Assays to Measure Effects of Regulatory Ligands on AMPK. in D Neumann & B Viollet (eds), AMPK : Methods and Protocols. Methods in Molecular Biology, vol. 1732, Humana Press, New York, pp. 69-86. https://doi.org/10.1007/978-1-4939-7598-3_5
Vara Ciruelos, D, Dandapani, M, Gray, A, Agbani, EO, Evans, AM & Hardie, DG 2018, 'Genotoxic Damage Activates the AMPK-α1 Isoform in the Nucleus via Ca2+/CaMKK2 Signaling to Enhance Tumor Cell Survival' Molecular Cancer Research, vol. 16, no. 2, pp. 345-357. https://doi.org/10.1158/1541-7786.MCR-17-0323
Hawley, SA, Fyffe, FA, Russell, FM, Gowans, GJ & Hardie, DG 2018, Intact Cell Assays to Monitor AMPK and Determine the Contribution of the AMP-Binding or ADaM Sites to Activation. in D Neumann & B Viollet (eds), AMPK : Methods and Protocols. Methods in Molecular Biology, vol. 1732, Humana Press, New York, pp. 239-253. https://doi.org/10.1007/978-1-4939-7598-3_16
Thomas, EC, Hook, SC, Gray, A, Chadt, A, Carling, D, Al-Hasani, H, Heesom, KJ, Hardie, DG & Tavaré, JM 2018, 'Isoform-specific AMPK association with TBC1D1 is reduced by a mutation associated with severe obesity' The Biochemical journal, vol. 475, no. 18, pp. 2969-2983. https://doi.org/10.1042/BCJ20180475
Hardie, DG 2018, 'Keeping the home fires burning: AMP-activated protein kinase' Journal of the Royal Society Interface, vol. 15, no. 138, 20170774. https://doi.org/10.1098/rsif.2017.0774
Moral-Sanz, J, Lewis, SA, MacMillan, S, Ross, FA, Thomson, A, Viollet, B, Foretz, M, Moran, C, Hardie, DG & Evans, AM 2018, 'The LKB1-AMPK-α1 signaling pathway triggers hypoxic pulmonary vasoconstriction downstream of mitochondria' Science Signaling, vol. 11, no. 550, eaau0296. https://doi.org/10.1126/scisignal.aau0296
Salt, IP & Hardie, DG 2017, 'AMP-Activated Protein Kinase: An Ubiquitous Signaling Pathway With Key Roles in the Cardiovascular System' Circulation Research, vol. 120, no. 11, pp. 1825-1841. https://doi.org/10.1161/CIRCRESAHA.117.309633
Hardie, DG & Lin, S-C 2017, 'AMP-activated protein kinase - not just an energy sensor' F1000 Research, vol. 6, 1724, pp. 1-11. https://doi.org/10.12688/f1000research.11960.1
Hardie, DG 2017, 'An Oncogenic Role for the Ubiquitin Ligase UBE2O by Targeting AMPK-α2 for Degradation' Cancer Cell, vol. 31, no. 2, pp. 163-165. https://doi.org/10.1016/j.ccell.2017.01.010
Xu, Y, Gray, A, Hardie, DG, Uzun, A, Shaw, S, Padbury, J, Phornphutkul, C & Tseng, Y-T 2017, 'A novel, de novo mutation in PRKAG2 gene: infantile-onset phenotype and signaling pathway involved' American Journal of Physiology: Heart and Circulatory Physiology, vol. 313, no. 2, pp. H283-H292. https://doi.org/10.1152/ajpheart.00813.2016
Lopez-Mejia, IC, Lagarrigue, S, Giralt, A, Martinez-Carreres, L, Zanou, N, Denechaud, P-D, Castillo-Armengol, J, Chavey, C, Orpinell, M, Delacuisine, B, Nasrallah, A, Collodet, C, Zhang, L, Viollet, B, Hardie, DG & Fajas, L 2017, 'CDK4 Phosphorylates AMPKα2 to Inhibit Its Activity and Repress Fatty Acid Oxidation' Molecular Cell, vol. 68, no. 2, pp. 336-349.e6. https://doi.org/10.1016/j.molcel.2017.09.034
Zhang, C-S, Hawley, SA, Zong, Y, Li, M, Wang, Z, Gray, A, Ma, T, Cui, J, Feng, J-W, Zhu, M, Wu, Y-Q, Li, TY, Ye, Z, Lin, S-Y, Yin, H, Piao, H-L, Hardie, DG & Lin, S-C 2017, 'Fructose-1,6-bisphosphate and aldolase mediate glucose sensing by AMPK' Nature, vol. 548, no. 7665, pp. 112-116. https://doi.org/10.1038/nature23275
Ross, FA, Hawley, SA, Auciello, FR, Gowans, GJ, Atrih, A, Lamont, DJ & Hardie, DG 2017, 'Mechanisms of Paradoxical Activation of AMPK by the Kinase Inhibitors SU6656 and Sorafenib' Cell Chemical Biology, vol. 24, no. 7, pp. 813-824.e4. https://doi.org/10.1016/j.chembiol.2017.05.021
Hardie, DG 2017, 'Targeting an energy sensor to treat diabetes' Science, vol. 357, no. 6350, pp. 455-456. https://doi.org/10.1126/science.aao1913
Rena, G, Hardie, DG & Pearson, ER 2017, 'The mechanisms of action of metformin' Diabetologia, vol. 60, no. 9, pp. 1577-1585. https://doi.org/10.1007/s00125-017-4342-z
Hawley, S. A., Ford, R. J., Smith, B. K., Gowans, G. J., Mancini, S. J., Pitt, R. D., Day, E. A., Salt, I. P., Steinberg, G. R. and Hardie, D. G. (2016) The Na+/Glucose Cotransporter Inhibitor Canagliflozin Activates AMPK by Inhibiting Mitochondrial Function and Increasing Cellular AMP Levels. Diabetes. 65, 2784-2794 d.o.i 10.2337/db16-0058 PMC: 27381369
Ross, F. A., Jensen, T. E. and Hardie, D. G. (2016) Differential regulation by AMP and ADP of AMPK complexes containing different gamma subunit isoforms. The Biochemical journal. 473, 189-199 d.o.i 10.1042/BJ20150910 Pubmed: 4700476 PMC: 26542978
Fogarty, S., Ross, F. A., Vara Ciruelos, D., Gray, A., Gowans, G. J. and Hardie, D. G. (2016) AMPK Causes Cell Cycle Arrest in LKB1-Deficient Cells via Activation of CAMKK2. Molecular cancer research : MCR. 14, 683-695 d.o.i 10.1158/1541-7786.MCR-15-0479 PMC: 27141100
Ross, F. A., MacKintosh, C. and Hardie, D. G. (2016) AMP-activated protein kinase: a cellular energy sensor that comes in twelve flavours. The FEBS journald.o.i 10.1111/febs.13698 PMC: 26934201
Hardie, D. G., Schaffer, B. E. and Brunet, A. (2016) AMPK: An Energy-Sensing Pathway with Multiple Inputs and Outputs. Trends in cell biology. 26, 190-201 d.o.i 10.1016/j.tcb.2015.10.013 PMC: 26616193
Hardie, D. G. (2015) Molecular Pathways: Is AMPK a Friend or a Foe in Cancer? Clinical cancer research : an official journal of the American Association for Cancer Research. 21, 3836-3840 d.o.i 10.1158/1078-0432.CCR-14-3300 Pubmed: 4558946 PMC: 26152739
Gowans, G. J., Hawley, S. A., Ross, F. A. and Hardie, D. G. (2013) AMP is a true physiological regulator of AMP-activated protein kinase by both allosteric activation and enhancing net phosphorylation. Cell metabolism. 18, 556-566 d.o.i 10.1016/j.cmet.2013.08.019 Pubmed: 3791399 PMC: 24093679
Hardie, D. G. and Alessi, D. R. (2013) LKB1 and AMPK and the cancer-metabolism link - ten years after. BMC biology. 11, 36 d.o.i 10.1186/1741-7007-11-36 Pubmed: 3626889 PMC: 23587167
O'Neill, L. A. and Hardie, D. G. (2013) Metabolism of inflammation limited by AMPK and pseudo-starvation. Nature. 493, 346-355 d.o.i 10.1038/nature11862 PMC: 23325217
Hawley, S. A., Fullerton, M. D., Ross, F. A., Schertzer, J. D., Chevtzoff, C., Walker, K. J., Peggie, M. W., Zibrova, D., Green, K. A., Mustard, K. J., Kemp, B. E., Sakamoto, K., Steinberg, G. R. and Hardie, D. G. (2012) The ancient drug salicylate directly activates AMP-activated protein kinase. Science. 336, 918-922 d.o.i 10.1126/science.1215327 pubmed: 3399766 PMC: 22517326