Ca2+/calmodulin-dependent protein kinase II (CaMKII) is an enzyme with important regulatory functions in the heart and brain, and its chronic activation can be pathological. CaMKII activation is seen in heart failure, and can directly induce pathological changes in ion channels, Ca2+ handling and gene transcription1. Here, in human, rat and mouse, we identify a novel mechanism linking CaMKII and hyperglycaemic signalling in diabetes mellitus, which is a key risk factor for heart2 and neurodegenerative diseases3,4. Acute hyperglycaemia causes covalent modification of CaMKII by O-linked N-acetylglucosamine (O-GlcNAc). O-GlcNAc modification of CaMKII at Ser 279 activates CaMKII autonomously, creating molecular memory even after Ca2+ concentration declines. O-GlcNAc-modified CaMKII is increased in the heart and brain of diabetic humans and rats. In cardiomyocytes, increased glucose concentration significantly enhances CaMKII-dependent activation of spontaneous sarcoplasmic reticulum Ca2+ release events that can contribute to cardiac mechanical dysfunction and arrhythmias1. These effects were prevented by pharmacological inhibition of O-GlcNAc signalling or genetic ablation of CaMKIIδ. In intact perfused hearts, arrhythmias were aggravated by increased glucose concentration through O-GlcNAc- and CaMKII-dependent pathways. In diabetic animals, acute blockade of O-GlcNAc inhibited arrhythmogenesis. Thus, O-GlcNAc modification of CaMKII is a novel signalling event in pathways that may contribute critically to cardiac and neuronal pathophysiology in diabetes and other diseases.
Subscribe to Journal
Get full journal access for 1 year
only $3.90 per issue
All prices are NET prices.
VAT will be added later in the checkout.
Tax calculation will be finalised during checkout.
Rent or Buy article
Get time limited or full article access on ReadCube.
All prices are NET prices.
Anderson, M. E., Brown, J. H. & Bers, D. M. CaMKII in myocardial hypertrophy and heart failure. J. Mol. Cell. Cardiol. 51, 468–473 (2011)
Roger, V. L. et al. Heart disease and stroke statistics—2011 update: a report from the American Heart Association. Circulation 123, e18–e209 (2011)
Haan, M. N. Therapy Insight: type 2 diabetes mellitus and the risk of late-onset Alzheimer’s disease. Nature Clin. Pract. Neurol. 2, 159–166 (2006)
Biessels, G. J., Staekenborg, S., Brunner, E., Brayne, C. & Scheltens, P. Risk of dementia in diabetes mellitus: a systematic review. Lancet Neurol. 5, 64–74 (2006)
Rosenberg, O. S., Deindl, S., Sung, R. J., Nairn, A. C. & Kuriyan, J. Structure of the autoinhibited kinase domain of CaMKII and SAXS analysis of the holoenzyme. Cell 123, 849–860 (2005)
Takao, K. et al. Visualization of synaptic Ca2+/calmodulin-dependent protein kinase II activity in living neurons. J. Neurosci. 25, 3107–3112 (2005)
Erickson, J. R., Patel, R., Ferguson, A., Bossuyt, J. & Bers, D. M. Fluorescence resonance energy transfer-based sensor Camui provides new insight into mechanisms of calcium/calmodulin-dependent protein kinase II activation in intact cardiomyocytes. Circ. Res. 109, 729–738 (2011)
Hudmon, A. & Schulman, H. Structure-–function of the multifunctional Ca2+/calmodulin-dependent protein kinase II. Biochem. J. 364, 593–611 (2002)
Erickson, J. R. et al. A dynamic pathway for calcium-independent activation of CaMKII by methionine oxidation. Cell 133, 462–474 (2008)
Hart, G. W., Housley, M. P. & Slawson, C. Cycling of O-linked β-N-acetylglucosamine on nucleocytoplasmic proteins. Nature 446, 1017–1022 (2007)
Chatham, J. C. & Marchase, R. B. The role of protein O-linked β-N-acetylglucosamine in mediating cardiac stress responses. Biochim Biophys Acta. 1800, 57–66 (2010)
Zhu-Mauldin, X., Marsh, S. A., Zou, L., Marchase, R. B. & Chatham, J. C. Modification of STIM1 by O-linked N-acetylglucosamine (O-GlcNAc) attenuates store-operated calcium entry in neonatal cardiomyocytes. J. Biol. Chem. 287, 39094–39106 (2012)
Rengifo, J., Gibson, C. J., Winkler, E., Collin, T. & Ehrlich, B. E. Regulation of the inositol 1,4,5-trisphosphate receptor type I by O-GlcNAc glycosylation. J. Neurosci. 27, 13813–13821 (2007)
Bimboese, P., Gibson, C. J., Schmidt, S., Xiang, W. & Ehrlich, B. E. Isoform-specific regulation of the inositol 1,4,5-trisphosphate receptor by O-linked glycosylation. J. Biol. Chem. 286, 15688–15697 (2011)
Dias, W. B., Cheung, W. D., Wang, Z. & Hart, G. W. Regulation of calcium/calmodulin-dependent kinase IV by O-GlcNAc modification. J. Biol. Chem. 284, 21327–21337 (2009)
Kreppel, L. K., Blomberg, M. A. & Hart, G. W. Dynamic glycosylation of nuclear and cytosolic proteins. Cloning and characterization of a unique O-GlcNAc transferase with multiple tetratricopeptide repeats. J. Biol. Chem. 272, 9308–9315 (1997)
Gao, Y., Wells, L., Comer, F. I., Parker, G. J. & Hart, G. W. Dynamic O-glycosylation of nuclear and cytosolic proteins: cloning and characterization of a neutral, cytosolic β-N-acetylglucosaminidase from human brain. J. Biol. Chem. 276, 9838–9845 (2001)
Hoch, B., Meyer, R., Hetzer, R., Krause, E. G. & Karczewski, P. Identification and expression of δ-isoforms of the multifunctional Ca2+/calmodulin-dependent protein kinase in failing and nonfailing human myocardium. Circ. Res. 84, 713–721 (1999)
Zhang, T. et al. The δC isoform of CaMKII is activated in cardiac hypertrophy and induces dilated cardiomyopathy and heart failure. Circ. Res. 92, 912–919 (2003)
Ling, H. et al. Requirement for Ca2+/calmodulin-dependent kinase II in the transition from pressure overload-induced cardiac hypertrophy to heart failure in mice. J. Clin. Invest. 119, 1230–1240 (2009)
Butler, A. E. et al. Diabetes due to a progressive defect in β-cell mass in rats transgenic for human islet amyloid polypeptide (HIP Rat): a new model for type 2 diabetes. Diabetes 53, 1509–1516 (2004)
Kohlhaas, M. et al. Increased sarcoplasmic reticulum calcium leak but unaltered contractility by acute CaMKII overexpression in isolated rabbit cardiac myocytes. Circ. Res. 98, 235–244 (2006)
Guo, T., Zhang, T., Mestril, R. & Bers, D. M. Ca2+/calmodulin-dependent protein kinase II phosphorylation of ryanodine receptor does affect calcium sparks in mouse ventricular myocytes. Circ. Res. 99, 398–406 (2006)
Zhao, Z. et al. Angiotensin II induces afterdepolarizations via reactive oxygen species and calmodulin kinase II signaling. J. Mol. Cell. Cardiol. 50, 128–136 (2011)
Del Gobbo, L. C. et al. Low serum magnesium concentrations are associated with a high prevalence of premature ventricular complexes in obese adults with type 2 diabetes. Cardiovasc. Diabetol. 11, 23 (2012)
Luo, M. et al. Diabetes increases mortality after myocardial infarction by oxidizing CaMKII. J. Clin. Invest. 123, 1262–1274 (2013)
Ashpole, N. M. & Hudmon, A. Excitotoxic neuroprotection and vulnerability with CaMKII inhibition. Mol. Cell. Neurosci. 46, 720–730 (2011)
Despa, S. et al. Hyperamylinemia contributes to cardiac dysfunction in obesity and diabetes: a study in humans and rats. Circ. Res. 110, 598–608 (2012)
van Oort, R. J. et al. Ryanodine receptor phosphorylation by calcium/calmodulin-dependent protein kinase II promotes life-threatening ventricular arrhythmias in mice with heart failure. Circulation 122, 2669–2679 (2010)
Curtis, M. J. & Walker, M. J. Quantification of arrhythmias using scoring systems: an examination of seven scores in an in vivo model of regional myocardial ischaemia. Cardiovasc. Res. 22, 656–665 (1988)
We thank Y. Hayashi for providing initial Camui samples, H. Schulman for helpful discussions, K. Margulies for human heart samples, L.-W. Jin, M. Melara and the University of California, Davis Alzheimer’s Disease Center (NIH-P30AG010129) for human brain samples, and J. H. Brown for providing CaMKIIδ-knockout mice. We thank Pfizer, Inc. for the gift of a breeding pair of HIP rats to F.D. This work was supported by American Heart Association 13SDG14680072 and National Institutes of Health (NIH) T32HL86350 (J.R.E.); NIH 1R01HL118474-01A1, NSF CBET 1133339, ADA 1-13-IN-70 and AHA 13GRNT16470034 (F.D.); NIH R01DK61671 and P01HL107153 (G.W.H.); NIH R01HL111600 (C.M.R.); NIH P01HL080101, R37HL30077 and Fondation Leducq Transatlantic CaMKII Alliance (D.M.B.). G.W.H. receives a share of royalty on sales of the CTD 110.6 antibody, which are managed by Johns Hopkins University.
The authors declare no competing financial interests.
Extended data figures and tables
Extended Data Figure 1 O-GlcNAc effect is not abolished by T286A or CM280/1VV mutation and CaMKII regulatory domain contains consensus O-GlcNAc modification sites.
a, Increased glucose concentration, but not osmolarity-matched mannitol, activates CaMKII in HEK cells (n = 9). b, O-GlcNAc-dependent CaMKII activation is reduced but still present in T286A-mutant Camui (n = 9). WT, wild type. c, Glucose-dependent CaMKII activation is preserved in CM280/281VV-mutant Camui expressed in HEK-cell lysates (n = 9). d, Activation of Camui by increased glucose is blunted in the S279A mutant and ablated entirely by DON (n values: wild type = 100, wild type + DON = 72, S279A = 57, S279A + DON = 44 cells). e, These sites are conserved in all known isoforms of CaMKII and in a wide variety of mammalian species. Data are mean ± s.e.m. * P < 0.05, **P < 0.01 versus control.
a, Immunoblot with an O-GlcNAc-specific antibody shows O-GlcNAc modification of CaMKII is inducible by increased glucose availability and is enhanced by Iso treatment (n values indicated). b, O-GlcNAc modification of CaMKII is reversed by β-elimination reaction before immunoblot. c, O-GlcNAc modification of CaMKII is ablated by DON and enhanced by Thm-G. d, Autophosphorylation of cardiac CaMKII is significantly increased in a rat model of diabetes. n = 3 for all immunoblots except where indicated. Data are mean ± s.e.m. *P < 0.05, **P < 0.01 versus control.
A synthetic peptide encoding the regulatory domain of CaMKII was subjected to in vitro O-GlcNAc labelling followed by ETD-MS analysis. Examination of the 507.25 m/z peptide fragment (top right inset) indicates the presence of an O-GlcNAc modification at S279 (c6 to c7 fragmentation).
a, b, Sarcoplasmic reticulum (SR) content is unaffected by Thm-G (a) or DON (b) in isolated rat myocytes (n values indicated). c, Mannitol does not enhance calcium spark frequency in isolated rat myocytes. d, e, Ca2+ transient amplitude (d, n = 13) and SR content (e, n = 13) are unaffected by Thm-G treatment in isolated myocytes from wild-type (WT) or CaMKIIδ-knockout mice. Data are mean ± s.e.m. NS, no significant difference.
a, b, Simultaneous treatment with 3,500 mg dl−1 glucose and Thm-G greatly enhances spark frequency (a) and SR Ca2+ depletion (b) in isolated rat myocytes (n = 6). Data are mean ± s.e.m.
Extended Data Figure 6 Diastolic calcium elevation under high glucose is suppressed by pre-treatment with 50 mM DON.
a, Average diastolic calcium elevation at baseline and following treatment with either high glucose (HG) or DON plus high glucose. Calcium elevation was measured as the percentage increase in the diastolic calcium signal relative to the amplitude of the following transient (n = 3). b, Example transients during baseline conditions (black) and after treatment with either high glucose or DON plus high glucose (grey). Minimal diastolic calcium elevation occurs after pre-treatment with DON. n = 3–4 rats for all data points. c, CaMKII activity is enhanced in heart lysate from diabetic rats (n = 3), and this effect is blunted by treatment with DON. Data are mean ± s.e.m. *P < 0.05 versus control.
About this article
Cite this article
Erickson, J., Pereira, L., Wang, L. et al. Diabetic hyperglycaemia activates CaMKII and arrhythmias by O-linked glycosylation. Nature 502, 372–376 (2013). https://doi.org/10.1038/nature12537
Cancer Cell International (2021)
Knockout of interleukin-17A diminishes ventricular arrhythmia susceptibility in diabetic mice via inhibiting NF-κB-mediated electrical remodeling
Acta Pharmacologica Sinica (2021)
Scientific Reports (2021)
Scientific Reports (2021)
Pflügers Archiv - European Journal of Physiology (2021)