Carbonylation induces heterogeneity in cardiac ryanodine receptor function in diabetes mellitus

Chun Hong Shao, Chengju Tian, Shouqiang Ouyang, Caronda J. Moore, Fadhel Alomar, Ina Nemet, Alicia D'Souza, Ryoji Nagai, Shelby Kutty, George J. Rozanski, Sasanka Ramanadham, Jaipaul Singh, Keshore R Bidasee

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Abstract

Heart failure and arrhythmias occur at 3 to 5 times higher rates among individuals with diabetes mellitus, compared with age-matched, healthy individuals. Studies attribute these defects in part to alterations in the function of cardiac type 2 ryanodine receptors (RyR2s), the principal Ca2+-release channels on the internal sarcoplasmic reticulum (SR). To date, mechanisms underlying RyR2 dysregulation in diabetes remain poorly defined. A rat model of type 1 diabetes, in combination with echocardiography, in vivo and ex vivo hemodynamic studies, confocal microscopy, Western blotting, mass spectrometry, site-directed mutagenesis, and [3H]ryanodine binding, lipid bilayer, and transfection assays, was used to determine whether post-translational modification by reactive carbonyl species (RCS) represented a contributing cause. After 8 weeks of diabetes, spontaneous Ca2+ release in ventricular myocytes increased ∼5-fold. Evoked Ca2+ release from the SR was nonuniform (dyssynchronous). Total RyR2 protein levels remained unchanged, but the ability to bind the Ca2+-dependent ligand [3H]ryanodine was significantly reduced. Western blotting and mass spectrometry revealed RCS adducts on select basic residues. Mutation of residues to delineate the physiochemical impact of carbonylation yielded channels with enhanced or reduced cytoplasmic Ca2+ responsiveness. The prototype RCS methylglyoxal increased and then decreased the RyR2 open probability. Methylglyoxal also increased spontaneous Ca2+ release and induced Ca2+ waves in healthy myocytes. Treatment of diabetic rats with RCS scavengers normalized spontaneous and evoked Ca2+ release from the SR, reduced carbonylation of RyR2s, and increased binding of [3H]ryanodine to RyR2s. From these data, we conclude that post-translational modification by RCS contributes to the heterogeneity in RyR2 activity that is seen in experimental diabetes.

Original languageEnglish (US)
Pages (from-to)383-399
Number of pages17
JournalMolecular pharmacology
Volume82
Issue number3
DOIs
StatePublished - Sep 1 2012

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Ryanodine Receptor Calcium Release Channel
Diabetes Mellitus
Ryanodine
Sarcoplasmic Reticulum
Pyruvaldehyde
Post Translational Protein Processing
Muscle Cells
Mass Spectrometry
Western Blotting
Lipid Bilayers
Site-Directed Mutagenesis
Type 1 Diabetes Mellitus
Confocal Microscopy
Transfection
Echocardiography
Cardiac Arrhythmias
Heart Failure
Hemodynamics
Ligands
Mutation

ASJC Scopus subject areas

  • Molecular Medicine
  • Pharmacology

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Carbonylation induces heterogeneity in cardiac ryanodine receptor function in diabetes mellitus. / Shao, Chun Hong; Tian, Chengju; Ouyang, Shouqiang; Moore, Caronda J.; Alomar, Fadhel; Nemet, Ina; D'Souza, Alicia; Nagai, Ryoji; Kutty, Shelby; Rozanski, George J.; Ramanadham, Sasanka; Singh, Jaipaul; Bidasee, Keshore R.

In: Molecular pharmacology, Vol. 82, No. 3, 01.09.2012, p. 383-399.

Research output: Contribution to journalArticle

Shao, CH, Tian, C, Ouyang, S, Moore, CJ, Alomar, F, Nemet, I, D'Souza, A, Nagai, R, Kutty, S, Rozanski, GJ, Ramanadham, S, Singh, J & Bidasee, KR 2012, 'Carbonylation induces heterogeneity in cardiac ryanodine receptor function in diabetes mellitus', Molecular pharmacology, vol. 82, no. 3, pp. 383-399. https://doi.org/10.1124/mol.112.078352
Shao, Chun Hong ; Tian, Chengju ; Ouyang, Shouqiang ; Moore, Caronda J. ; Alomar, Fadhel ; Nemet, Ina ; D'Souza, Alicia ; Nagai, Ryoji ; Kutty, Shelby ; Rozanski, George J. ; Ramanadham, Sasanka ; Singh, Jaipaul ; Bidasee, Keshore R. / Carbonylation induces heterogeneity in cardiac ryanodine receptor function in diabetes mellitus. In: Molecular pharmacology. 2012 ; Vol. 82, No. 3. pp. 383-399.
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