S-Nitrosylation of Sarcomeric Proteins Depresses Myofilament Ca2+)Sensitivity in Intact Cardiomyocytes.

Autor:	Figueiredo-Freitas C; 1 Department of Biomedical Sciences, College of Medicine, Florida State University , Tallahassee, Florida.; 2 Instituto de Bioquímica Médica Leopoldo de Meis (IBqM), Federal University of Rio de Janeiro , Rio de Janeiro, Brazil .; 3 Department of Molecular and Cellular Pharmacology, Miller School of Medicine, University of Miami , Miami, Florida., Dulce RA; 4 Interdisciplinary Stem Cell Institute, University of Miami , Miami, Florida., Foster MW; 5 Pulmonary, Allergy and Critical Care Medicine, Duke University Medical Center , Durham, North Carolina.; 6 Proteomics and Metabolomics Shared Resource, Duke University Medical Center , Durham, North Carolina., Liang J; 3 Department of Molecular and Cellular Pharmacology, Miller School of Medicine, University of Miami , Miami, Florida., Yamashita AM; 2 Instituto de Bioquímica Médica Leopoldo de Meis (IBqM), Federal University of Rio de Janeiro , Rio de Janeiro, Brazil ., Lima-Rosa FL; 2 Instituto de Bioquímica Médica Leopoldo de Meis (IBqM), Federal University of Rio de Janeiro , Rio de Janeiro, Brazil ., Thompson JW; 6 Proteomics and Metabolomics Shared Resource, Duke University Medical Center , Durham, North Carolina., Moseley MA; 6 Proteomics and Metabolomics Shared Resource, Duke University Medical Center , Durham, North Carolina., Hare JM; 4 Interdisciplinary Stem Cell Institute, University of Miami , Miami, Florida., Nogueira L; 2 Instituto de Bioquímica Médica Leopoldo de Meis (IBqM), Federal University of Rio de Janeiro , Rio de Janeiro, Brazil ., Sorenson MM; 2 Instituto de Bioquímica Médica Leopoldo de Meis (IBqM), Federal University of Rio de Janeiro , Rio de Janeiro, Brazil ., Pinto JR; 1 Department of Biomedical Sciences, College of Medicine, Florida State University , Tallahassee, Florida.; 3 Department of Molecular and Cellular Pharmacology, Miller School of Medicine, University of Miami , Miami, Florida.
Jazyk:	angličtina
Zdroj:	Antioxidants & redox signaling [Antioxid Redox Signal] 2015 Nov 01; Vol. 23 (13), pp. 1017-34.
DOI:	10.1089/ars.2015.6275
Abstrakt:	Aims: The heart responds to physiological and pathophysiological stress factors by increasing its production of nitric oxide (NO), which reacts with intracellular glutathione to form S-nitrosoglutathione (GSNO), a protein S-nitrosylating agent. Although S-nitrosylation protects some cardiac proteins against oxidative stress, direct effects on myofilament performance are unknown. We hypothesize that S-nitrosylation of sarcomeric proteins will modulate the performance of cardiac myofilaments. Results: Incubation of intact mouse cardiomyocytes with S-nitrosocysteine (CysNO, a cell-permeable low-molecular-weight nitrosothiol) significantly decreased myofilament Ca(2+) sensitivity. In demembranated (skinned) fibers, S-nitrosylation with 1 μM GSNO also decreased Ca(2+) sensitivity of contraction and 10 μM reduced maximal isometric force, while inhibition of relaxation and myofibrillar ATPase required higher concentrations (≥ 100 μM). Reducing S-nitrosylation with ascorbate partially reversed the effects on Ca(2+) sensitivity and ATPase activity. In live cardiomyocytes treated with CysNO, resin-assisted capture of S-nitrosylated protein thiols was combined with label-free liquid chromatography-tandem mass spectrometry to quantify S-nitrosylation and determine the susceptible cysteine sites on myosin, actin, myosin-binding protein C, troponin C and I, tropomyosin, and titin. The ability of sarcomere proteins to form S-NO from 10-500 μM CysNO in intact cardiomyocytes was further determined by immunoblot, with actin, myosin, myosin-binding protein C, and troponin C being the more susceptible sarcomeric proteins. Innovation and Conclusions: Thus, specific physiological effects are associated with S-nitrosylation of a limited number of cysteine residues in sarcomeric proteins, which also offer potential targets for interventions in pathophysiological situations.
Databáze:	MEDLINE
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