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

Title: S-Nitrosylation of Sarcomeric Proteins Depresses Myofilament Ca2+)Sensitivity in Intact Cardiomyocytes.
Name(s): Figueiredo-Freitas, Cícero, author
Dulce, Raul A, author
Foster, Matthew W, author
Liang, Jingsheng, author
Yamashita, Aline M S, author
Lima-Rosa, Frederico L, author
Thompson, J Will, author
Moseley, M Arthur, author
Hare, Joshua M, author
Nogueira, Leonardo, author
Sorenson, Martha M, author
Pinto, José Renato, author
Type of Resource: text
Genre: Journal Article
Date Issued: 2015-11-01
Physical Form: computer
online resource
Extent: 1 online resource
Language(s): English
Abstract/Description: 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. 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. 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.
Identifier: FSU_pmch_26421519 (IID), 10.1089/ars.2015.6275 (DOI), PMC4649751 (PMCID), 26421519 (RID), 26421519 (EID)
Grant Number: HL103840, 5R01 HL094849, HL106121, K99 HL103840, R00 HL103840
Publication Note: This NIH-funded author manuscript originally appeared in PubMed Central at
Subject(s): Animals
Ca(2+) Mg(2+)-ATPase/metabolism
Calcium Signaling
Cells, Cultured
Cysteine/analogs & derivatives
Mice, 129 Strain
Mice, Inbred C57BL
Muscle Proteins/metabolism
Myocardial Contraction
Myocytes, Cardiac/metabolism
Nitric Oxide/metabolism
Oxidative Stress
Persistent Link to This Record:
Owner Institution: FSU
Is Part Of: Antioxidants & redox signaling.
Issue: iss. 13, vol. 23

Choose the citation style.
Figueiredo-Freitas, C., Dulce, R. A., Foster, M. W., Liang, J., Yamashita, A. M. S., Lima-Rosa, F. L., … Pinto, J. R. (2015). S-Nitrosylation of Sarcomeric Proteins Depresses Myofilament Ca2+)Sensitivity in Intact Cardiomyocytes. Antioxidants & Redox Signaling. Retrieved from