Quantitative In Vivo Proteomics of Metformin Response in Liver Reveals AMPK-Dependent and -Independent Signaling Networks

Overview

Journal Cell Rep

Publisher Cell Press

Specialties Biology
Cell Biology
Molecular Biology

Date 2019 Dec 5

PMID 31801093

Citations 20

Authors

Benjamin D Stein

Diego Calzolari

Kristina Hellberg

Ying S Hu

Lin He

Chien-Min Hung

Erin Q Toyama

Debbie S Ross

Bjorn F Lillemeier

Lewis C Cantley

John R Yates 3rd

Reuben J Shaw

Affiliations

Soon will be listed here.

Abstract

Metformin is the front-line treatment for type 2 diabetes worldwide. It acts via effects on glucose and lipid metabolism in metabolic tissues, leading to enhanced insulin sensitivity. Despite significant effort, the molecular basis for metformin response remains poorly understood, with a limited number of specific biochemical pathways studied to date. To broaden our understanding of hepatic metformin response, we combine phospho-protein enrichment in tissue from genetically engineered mice with a quantitative proteomics platform to enable the discovery and quantification of basophilic kinase substrates in vivo. We define proteins whose binding to 14-3-3 are acutely regulated by metformin treatment and/or loss of the serine/threonine kinase, LKB1. Inducible binding of 250 proteins following metformin treatment is observed, 44% of which proteins bind in a manner requiring LKB1. Beyond AMPK, metformin activates protein kinase D and MAPKAPK2 in an LKB1-independent manner, revealing additional kinases that may mediate aspects of metformin response. Deeper analysis uncovered substrates of AMPK in endocytosis and calcium homeostasis.

Citing Articles

AMPK Activation Downregulates TXNIP, Rab5, and Rab7 Within Minutes, Thereby Inhibiting the Endocytosis-Mediated Entry of Human Pathogenic Viruses.

Diesendorf V, La Rocca V, Teutsch M, Alattar H, Obernolte H, Kenst K Cells. 2025; 14(5).

PMID: 40072063 PMC: 11899703. DOI: 10.3390/cells14050334.

The Research Progress of Metformin Regulation of Metabolic Reprogramming in Malignant Tumors.

Sui Q, Yang H, Hu Z, Jin X, Chen Z, Jiang W Pharm Res. 2024; 41(11):2143-2159.

PMID: 39455505 DOI: 10.1007/s11095-024-03783-2.

Research progress on the role of mitochondria in the process of hepatic ischemia-reperfusion injury.

Zhou Y, Qiu T, Wang T, Yu B, Xia K, Guo J Gastroenterol Rep (Oxf). 2024; 12:goae066.

PMID: 38912038 PMC: 11193119. DOI: 10.1093/gastro/goae066.

Systematic Investigation of Dose-Dependent Protein Thermal Stability Changes to Uncover the Mechanisms of the Pleiotropic Effects of Metformin.

Yin K, Wu R ACS Pharmacol Transl Sci. 2024; 7(2):467-477.

PMID: 38357277 PMC: 10863438. DOI: 10.1021/acsptsci.3c00298.

Role of STIM1 in the Regulation of Cardiac Energy Substrate Preference.

Liu P, Yang Z, Wang Y, Sun A Int J Mol Sci. 2023; 24(17).

PMID: 37685995 PMC: 10487555. DOI: 10.3390/ijms241713188.

References

Cobbaut M, Van Lint J . Function and Regulation of Protein Kinase D in Oxidative Stress: A Tale of Isoforms. Oxid Med Cell Longev. 2018; 2018:2138502. PMC: 5944262. DOI: 10.1155/2018/2138502. View

Chauhan A, Liu X, Jing J, Lee H, Yadav R, Liu J . STIM2 interacts with AMPK and regulates calcium-induced AMPK activation. FASEB J. 2018; 33(2):2957-2970. PMC: 6338636. DOI: 10.1096/fj.201801225R. View

Kobayashi T, Hino S, Oue N, Asahara T, Zollo M, Yasui W . Glycogen synthase kinase 3 and h-prune regulate cell migration by modulating focal adhesions. Mol Cell Biol. 2006; 26(3):898-911. PMC: 1347031. DOI: 10.1128/MCB.26.3.898-911.2006. View

Chen S, Synowsky S, Tinti M, Mackintosh C . The capture of phosphoproteins by 14-3-3 proteins mediates actions of insulin. Trends Endocrinol Metab. 2011; 22(11):429-36. DOI: 10.1016/j.tem.2011.07.005. View

Reinhardt H, Yaffe M . Kinases that control the cell cycle in response to DNA damage: Chk1, Chk2, and MK2. Curr Opin Cell Biol. 2009; 21(2):245-55. PMC: 2699687. DOI: 10.1016/j.ceb.2009.01.018. View

Christoforides C, Rainero E, Brown K, Norman J, Toker A . PKD controls αvβ3 integrin recycling and tumor cell invasive migration through its substrate Rabaptin-5. Dev Cell. 2012; 23(3):560-72. PMC: 3443396. DOI: 10.1016/j.devcel.2012.08.008. View

Nelson M, Parker B, Burchfield J, Hoffman N, Needham E, Cooke K . Phosphoproteomics reveals conserved exercise-stimulated signaling and AMPK regulation of store-operated calcium entry. EMBO J. 2019; 38(24):e102578. PMC: 6912027. DOI: 10.15252/embj.2019102578. View

Yano T, Matsui T, Tamura A, Uji M, Tsukita S . The association of microtubules with tight junctions is promoted by cingulin phosphorylation by AMPK. J Cell Biol. 2014; 203(4):605-14. PMC: 3840929. DOI: 10.1083/jcb.201304194. View

Foretz M, Hebrard S, Leclerc J, Zarrinpashneh E, Soty M, Mithieux G . Metformin inhibits hepatic gluconeogenesis in mice independently of the LKB1/AMPK pathway via a decrease in hepatic energy state. J Clin Invest. 2010; 120(7):2355-69. PMC: 2898585. DOI: 10.1172/JCI40671. View

10.

Batalha I, Lowe C, Roque A . Platforms for enrichment of phosphorylated proteins and peptides in proteomics. Trends Biotechnol. 2011; 30(2):100-10. DOI: 10.1016/j.tibtech.2011.07.004. View

11.

Laplante M, Sabatini D . mTOR signaling in growth control and disease. Cell. 2012; 149(2):274-93. PMC: 3331679. DOI: 10.1016/j.cell.2012.03.017. View

12.

Cai S, Tee A, Short J, Bergeron J, Kim J, Shen J . Activity of TSC2 is inhibited by AKT-mediated phosphorylation and membrane partitioning. J Cell Biol. 2006; 173(2):279-89. PMC: 2063818. DOI: 10.1083/jcb.200507119. View

13.

Patel K, Foretz M, Marion A, Campbell D, Gourlay R, Boudaba N . The LKB1-salt-inducible kinase pathway functions as a key gluconeogenic suppressor in the liver. Nat Commun. 2014; 5:4535. PMC: 4143937. DOI: 10.1038/ncomms5535. View

14.

Christoforidis S, Miaczynska M, Ashman K, Wilm M, Zhao L, Yip S . Phosphatidylinositol-3-OH kinases are Rab5 effectors. Nat Cell Biol. 1999; 1(4):249-52. DOI: 10.1038/12075. View

15.

Dale S, Wilson W, EDELMAN A, Hardie D . Similar substrate recognition motifs for mammalian AMP-activated protein kinase, higher plant HMG-CoA reductase kinase-A, yeast SNF1, and mammalian calmodulin-dependent protein kinase I. FEBS Lett. 1995; 361(2-3):191-5. DOI: 10.1016/0014-5793(95)00172-6. View

16.

Russell R, Yuan H, Guan K . Autophagy regulation by nutrient signaling. Cell Res. 2013; 24(1):42-57. PMC: 3879708. DOI: 10.1038/cr.2013.166. View

17.

Cobbaut M, Derua R, Doppler H, Lou H, Vandoninck S, Storz P . Differential regulation of PKD isoforms in oxidative stress conditions through phosphorylation of a conserved Tyr in the P+1 loop. Sci Rep. 2017; 7(1):887. PMC: 5430542. DOI: 10.1038/s41598-017-00800-w. View

18.

Itoh Y, Sanosaka M, Fuchino H, Yahara Y, Kumagai A, Takemoto D . Salt-inducible Kinase 3 Signaling Is Important for the Gluconeogenic Programs in Mouse Hepatocytes. J Biol Chem. 2015; 290(29):17879-17893. PMC: 4505037. DOI: 10.1074/jbc.M115.640821. View

19.

Tian L, Chen J, Chen M, Gui C, Zhong C, Hong L . The p38 pathway regulates oxidative stress tolerance by phosphorylation of mitochondrial protein IscU. J Biol Chem. 2014; 289(46):31856-31865. PMC: 4231663. DOI: 10.1074/jbc.M114.589093. View

20.

Yaffe M . How do 14-3-3 proteins work?-- Gatekeeper phosphorylation and the molecular anvil hypothesis. FEBS Lett. 2002; 513(1):53-7. DOI: 10.1016/s0014-5793(01)03288-4. View