The major energy source for most cells is glucose, from which ATP is generated via glycolysis and/or oxidative metabolism. Glucose deprivation activates AMP-activated protein kinase (AMPK)1, but it is unclear whether this activation occurs solely via changes in AMP or ADP, the classical activators of AMPK2,3,4,5. Here, we describe an AMP/ADP-independent mechanism that triggers AMPK activation by sensing the absence of fructose-1,6-bisphosphate (FBP), with AMPK being progressively activated as extracellular glucose and intracellular FBP decrease. When unoccupied by FBP, aldolases promote the formation of a lysosomal complex containing at least v-ATPase, ragulator, axin, liver kinase B1 (LKB1) and AMPK, which has previously been shown to be required for AMPK activation6,7. Knockdown of aldolases activates AMPK even in cells with abundant glucose, whereas the catalysis-defective D34S aldolase mutant, which still binds FBP, blocks AMPK activation. Cell-free reconstitution assays show that addition of FBP disrupts the association of axin and LKB1 with v-ATPase and ragulator. Importantly, in some cell types AMP/ATP and ADP/ATP ratios remain unchanged during acute glucose starvation, and intact AMP-binding sites on AMPK are not required for AMPK activation. These results establish that aldolase, as well as being a glycolytic enzyme, is a sensor of glucose availability that regulates AMPK.

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We thank all other members of the S.-C.L. laboratory for suggestions and technical assistance, and the proteomics team at the University of Dundee (D. Lamont, A. Atrih, W. Chen and K. Beattie) for LC–MS analyses of nucleotides. D.G.H. was supported by an Investigator Award from the Wellcome Trust (097726) and a Programme Grant from Cancer Research UK (C37030/A15101). S.-C.L. was supported by grants from the National Key Research and Development Project of China (2016YFA0502001) and the National Natural Science Foundation of China (#31430094, #31690101, #31571214, #31601152 and #J1310027).

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Author notes

    • Chen-Song Zhang
    • , Simon A. Hawley
    • , Yue Zong
    •  & Mengqi Li

    These authors contributed equally to this work.


  1. State Key Laboratory for Cellular Stress Biology, Innovation Center for Cell Signaling Network, School of Life Sciences, Xiamen University, Fujian 361102, China

    • Chen-Song Zhang
    • , Yue Zong
    • , Mengqi Li
    • , Teng Ma
    • , Jiwen Cui
    • , Jin-Wei Feng
    • , Yu-Qing Wu
    • , Terytty Yang Li
    • , Zhiyun Ye
    • , Shu-Yong Lin
    •  & Sheng-Cai Lin
  2. Division of Cell Signalling and Immunology, College of Life Sciences, University of Dundee, Dundee DD1 5EH, UK

    • Simon A. Hawley
    • , Alexander Gray
    •  & D. Grahame Hardie
  3. Scientific Research Center for Translational Medicine, Dalian Institute of Chemical Physics, Chinese Academy of Sciences, Liaoning 116023, China

    • Zhichao Wang
    •  & Hai-Long Piao
  4. Key Laboratory of Separation Science for Analytical Chemistry, Dalian Institute of Chemical Physics, Chinese Academy of Sciences, Liaoning 116023, China

    • Zhichao Wang
  5. University of Chinese Academy of Sciences, Beijing 100049, China

    • Zhichao Wang
  6. Key Laboratory of Food Safety Research, Institute for Nutritional Sciences (INS), Shanghai Institutes for Biological Sciences (SIBS), Chinese Academy of Sciences (CAS), Shanghai 200031, China

    • Mingjiang Zhu
    •  & Huiyong Yin


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C.-S.Z., S.A.H., Y.Z., M.L., D.G.H. and S.-C.L. conceived the study. C.-S.Z., S.A.H., Y.Z. and M.L. performed most experiments with assistance from T.M., J.C., J.-W.F., A.G., Y.-Q.W., T.Y.L. and S.-Y.L. Y.Z. and Z.W. performed the CE–MS-based analysis of metabolites, and S.A.H., M.L. and M.Z. performed the analysis of adenylates. Z.Y., S.-Y.L., H.Y. and H.-L.P. helped with discussion and interpretation of results. D.G.H. and S.-C.L. wrote the manuscript.

Competing interests

The authors declare no competing financial interests.

Corresponding authors

Correspondence to D. Grahame Hardie or Sheng-Cai Lin.

Reviewer Information Nature thanks D. Tolan and the other anonymous reviewer(s) for their contribution to the peer review of this work.

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