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Cardioprotection by S-nitrosation of a cysteine switch on mitochondrial complex I

Abstract

Oxidative damage from elevated production of reactive oxygen species (ROS) contributes to ischemia-reperfusion injury in myocardial infarction and stroke. The mechanism by which the increase in ROS occurs is not known, and it is unclear how this increase can be prevented. A wide variety of nitric oxide donors and S-nitrosating agents protect the ischemic myocardium from infarction, but the responsible mechanisms are unclear1,2,3,4,5,6. Here we used a mitochondria-selective S-nitrosating agent, MitoSNO, to determine how mitochondrial S-nitrosation at the reperfusion phase of myocardial infarction is cardioprotective in vivo in mice. We found that protection is due to the S-nitrosation of mitochondrial complex I, which is the entry point for electrons from NADH into the respiratory chain. Reversible S-nitrosation of complex I slows the reactivation of mitochondria during the crucial first minutes of the reperfusion of ischemic tissue, thereby decreasing ROS production, oxidative damage and tissue necrosis. Inhibition of complex I is afforded by the selective S-nitrosation of Cys39 on the ND3 subunit, which becomes susceptible to modification only after ischemia. Our results identify rapid complex I reactivation as a central pathological feature of ischemia-reperfusion injury and show that preventing this reactivation by modification of a cysteine switch is a robust cardioprotective mechanism and hence a rational therapeutic strategy.

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Figure 1: S-nitrosation of mitochondrial proteins is required for S-nitrosation–mediated protection from cardiac ischemia-reperfusion injury.
Figure 2: Respiratory complex I ND3 Cys39 S-nitrosation is dependent on low complex I activity.
Figure 3: ND3 Cys39 S-nitrosation mediates inhibition of complex I activity and ROS production.
Figure 4: S-nitrosation of ND3 Cys39 underlies protection from ischemia-reperfusion injury by mitochondrial S-nitrosation in vivo and represents a general mechanism for cardioprotection.

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Acknowledgements

This study was supported by the UK Medical Research Council and grants from the UK Biotechnology and Biological Sciences Research Council (BB/I012923 to M.P.M. and R.C.H.), the Gates Cambridge Trust and the Canadian Institutes of Health Research (doctoral scholarship and postdoctoral fellowship to E.T.C.), the British Heart Foundation (PG/12/42/29655 to T.K.), the US National Institutes of Health (R01-HL071158 to P.S.B.) and the International Society for Heart Research (ISHR-ES/SERVIER research fellowship to C.M.). We thank L. Sazanov and J. Hirst for helpful discussions.

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E.T.C. designed research, carried out mass spectrometry, fluorescence and biochemical experiments, analyzed data from in vivo experiments and cowrote the paper. C.M., S.M.N. and V.R.P. carried out and analyzed data from in vivo experiments. J.R. assisted with DIGE experiments. A.L. and S.D. carried out mass spectrometry experiments. A.M.J. and H.M.C. assisted with research design and data interpretation. A.J.R. designed and performed bioinformatic experiments. L.P., I.M.F., R.C.H., K.S.L., R.A.J.S., T.K. and P.S.B. assisted with research design. M.P.M. designed and directed the project and cowrote the paper.

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Correspondence to Michael P Murphy.

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M.P.M. and R.A.J.S. hold an EU patent on the technology described in this publication.

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Chouchani, E., Methner, C., Nadtochiy, S. et al. Cardioprotection by S-nitrosation of a cysteine switch on mitochondrial complex I. Nat Med 19, 753–759 (2013). https://doi.org/10.1038/nm.3212

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