MICAL1 constrains cardiac stress responses and protects against disease by oxidizing CaMKII
Klitos Konstantinidis, … , Rodney L. Levine, Mark E. Anderson
Klitos Konstantinidis, … , Rodney L. Levine, Mark E. Anderson
Published August 4, 2020
Citation Information: J Clin Invest. 2020;130(9):4663-4678. https://doi.org/10.1172/JCI133181.
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Research Article Cardiology Cell biology

MICAL1 constrains cardiac stress responses and protects against disease by oxidizing CaMKII

Abstract

Oxidant stress can contribute to health and disease. Here we show that invertebrates and vertebrates share a common stereospecific redox pathway that protects against pathological responses to stress, at the cost of reduced physiological performance, by constraining Ca2+/calmodulin-dependent protein kinase II (CaMKII) activity. MICAL1, a methionine monooxygenase thought to exclusively target actin, and MSRB, a methionine reductase, control the stereospecific redox status of M308, a highly conserved residue in the calmodulin-binding (CaM-binding) domain of CaMKII. Oxidized or mutant M308 (M308V) decreased CaM binding and CaMKII activity, while absence of MICAL1 in mice caused cardiac arrhythmias and premature death due to CaMKII hyperactivation. Mimicking the effects of M308 oxidation decreased fight-or-flight responses in mice, strikingly impaired heart function in Drosophila melanogaster, and caused disease protection in human induced pluripotent stem cell–derived cardiomyocytes with catecholaminergic polymorphic ventricular tachycardia, a CaMKII-sensitive genetic arrhythmia syndrome. Our studies identify a stereospecific redox pathway that regulates cardiac physiological and pathological responses to stress across species.

Authors

Klitos Konstantinidis, Vassilios J. Bezzerides, Lo Lai, Holly M. Isbell, An-Chi Wei, Yuejin Wu, Meera C. Viswanathan, Ian D. Blum, Jonathan M. Granger, Danielle Heims-Waldron, Donghui Zhang, Elizabeth D. Luczak, Kevin R. Murphy, Fujian Lu, Daniel H. Gratz, Bruno Manta, Qiang Wang, Qinchuan Wang, Alex L. Kolodkin, Vadim N. Gladyshev, Thomas J. Hund, William T. Pu, Mark N. Wu, Anthony Cammarato, Mario A. Bianchet, Madeline A. Shea, Rodney L. Levine, Mark E. Anderson

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Figure 3

Met308 is critical for CaM binding and CaMKII activity.

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Met308 is critical for CaM binding and CaMKII activity. (A) Model interf...
(A) Model interfaces between (Ca2+)4-CaM residues E84, I85, A88, and M145 and CaMKII residues M308 (WT), M308-SO, M308V, or M308Q shown from 2 viewpoints (rotated by 90° about the vertical y axis). CaM side chains are red sticks, with E84 carboxyl oxygens (OE1, OE2) highlighted in pink, M145 sulfur (SD) atom shown as a yellow-orange sphere, CaMKII WT M308 (black), mutant M308-SO (green), M308V (burgundy), and M308Q (cyan) heteroatoms are shown as spheres; C-α carbon atoms (gray), hydrogen atoms (white), M308 and M308-SO, sulfur (yellow-orange), side chain oxygen of M308-SO (forest green). Models were aligned based on the backbone of CaMKII residues 294–311 (gray cylinder). (B) Fluorescence anisotropy measurements show binding of WT CaMKII CaM-binding-domain peptide compared with peptides with M308-SO, M308V, or M308Q at various (n = 9–11) CaM concentrations (WT and M308-SO data are the same as shown in Figure 1F). The curves represent nonlinear regression fit derived from the experimental data. Each data point represents the average of n = 2 measurements at each CaM concentration. (C) Fluorescence anisotropy measurements of the same peptides as B at baseline and after the addition of 50 μM Ca2+. Data represented as the mean of n = 3 replicates ± SEM. (D) Modeling of WT CaMKII holoenzyme activity compared with M308-SO CaMKII and M308V CaMKII with increasing frequency stimulation. Note that M308-SO and M308V curves are superimposed. (E) CaMKII kinase activity for WT and M308V recombinant proteins using syntide as a substrate with radioactive 32P-ATP. Data represented as the mean of n = 3 replicates. Bars denote mean ± SEM. (F) CaMKII kinase activity for WT and M308V recombinant proteins using syntide as a substrate with nonradioactive ATP using HPLC (representative image of n = 2 replicates). mAU, milliabsorbance units. **P < 0.01; ***P < 0.001 by 1-way ANOVA with Tukey’s multiple-comparisons test (C) or 2-tailed Student’s t test (E).