Abstract
Rationale: In the myocardium, redox/cysteine modification of proteins regulating Ca cycling can affect contraction and may have therapeutic value. Nitroxyl (HNO), the one-electron-reduced form of nitric oxide, enhances cardiac function in a manner that suggests reversible cysteine modifications of the contractile machinery. Objective:: To determine the effects of HNO modification in cardiac myofilament proteins. Methods and Results: The HNO-donor, 1-nitrosocyclohexyl acetate, was found to act directly on the myofilament proteins, increasing maximum force (Fmax) and reducing the concentration of Ca for 50% activation (Ca50) in intact and skinned cardiac muscles. The effects of 1-nitrosocyclohexyl acetate are reversible by reducing agents and distinct from those of another HNO donor, Angeli salt, which was previously reported to increase Fmax without affecting Ca50. Using a new mass spectrometry capture technique based on the biotin switch assay, we identified and characterized the formation by HNO of a disulfide-linked actin-tropomyosin and myosin heavy chain-myosin light chain 1. Comparison of the 1-nitrosocyclohexyl acetate and Angeli salt effects with the modifications induced by each donor indicated the actin-tropomyosin and myosin heavy chain-myosin light chain 1 interactions independently correlated with increased Ca sensitivity and force generation, respectively. Conclusions:: HNO exerts a direct effect on cardiac myofilament proteins increasing myofilament Ca responsiveness by promoting disulfide bond formation between critical cysteine residues. These findings indicate a novel, redox-based modulation of the contractile apparatus, which positively impacts myocardial function, providing further mechanistic insight for HNO as a therapeutic agent.
Original language | English (US) |
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Pages (from-to) | 1002-1011 |
Number of pages | 10 |
Journal | Circulation research |
Volume | 111 |
Issue number | 8 |
DOIs | |
State | Published - Sep 2012 |
Externally published | Yes |
Keywords
- calcium
- contractility
- nitroxyl
- oxidant signaling
- oxidation
- redox
- redox switch
ASJC Scopus subject areas
- Physiology
- Cardiology and Cardiovascular Medicine