S -nitrosylation and S -glutathionylation of Cys134 on troponin I have opposing competitive actions on Ca 2+ sensitivity in rat fast-twitch muscle fibers
Abstract
Nitric oxide is generated in skeletal muscle with activity and decreases Ca 2+ sensitivity of the contractile apparatus, putatively by S-nitrosylation of an unidentified protein. We investigated the mechanistic basis of this effect and its relationship to the oxidation-induced increase in Ca 2+ sensitivity in mammalian fast-twitch (FT) fibers mediated by S-glutathionylation of Cys134 on fast troponin I (TnI f ). Force-[Ca 2+ ] characteristics of the contractile apparatus in mechanically skinned fibers were assessed by direct activation with heavily Ca 2+ -buffered solutions. Treatment with S-nitrosylating agents, S-nitrosoglutathione (GSNO) or S-nitroso- N-acetyl-penicillamine (SNAP), decreased pCa 50 ( = −log 10 [Ca 2+ ] at half-maximal activation) by ~−0.07 pCa units in rat and human FT fibers without affecting maximum force, but had no effect on rat and human slow-twitch fibers or toad or chicken FT fibers, which all lack Cys134. The Ca 2+ sensitivity decrease was 1) fully reversed with dithiothreitol or reduced glutathione, 2) at least partially reversed with ascorbate, indicative of involvement of S-nitrosylation, and 3) irreversibly blocked by low concentration of the alkylating agent, N-ethylmaleimide (NEM). The biotin-switch assay showed that both GSNO and SNAP treatments caused S-nitrosylation of TnI f . S-glutathionylation pretreatment blocked the effects of S-nitrosylation on Ca 2+ sensitivity, and vice-versa. S-nitrosylation pretreatment prevented NEM from irreversibly blocking S-glutathionylation of TnI f and its effects on Ca 2+ sensitivity, and likewise S-glutathionylation pretreatment prevented NEM block of S-nitrosylation. Following substitution of TnI f into rat slow-twitch fibers, S-nitrosylation treatment caused decreased Ca 2+ sensitivity. These findings demonstrate that S-nitrosylation and S-glutathionylation exert opposing effects on Ca 2+ sensitivity in mammalian FT muscle fibers, mediated by competitive actions on Cys134 of TnI f .
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Various contractile-related proteins can undergo reversible post-translation modifications at cysteine residues on thiols, such as myosin, troponin, SERCA and ryanodine receptors, and these modifications are likely dependent on interactions between NO, reactive oxygen species and glutathione bioavailability.
“The potential influence of S-nitrosylation on excitation–contraction coupling is complex given that various contractile-related proteins can undergo reversible post-translation modifications at cysteine residues on thiols, such as myosin [140], troponin [141], SERCA [142] and ryanodine receptors (RyRs) [143,144], and that these post-translation protein modifications are likely dependent on inter”
From Effects of Dietary Nitrate Supplementation on Weightlifting Exercise Performance in Healthy Adults: A Systematic Review (2020) · cites this paper as [141]
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- Authors
- Dutka, T. L., Mollica, J. P., Lamboley, C. R., Weerakkody, V. C., Greening, D. W., Posterino, G. S., Murphy, R. M., & Lamb, G. D.
- Affiliations
- La Trobe University (AU)
- Corresponding author
- Lamb, G. D.
- Venue
- American Journal of Physiology-Cell Physiology
- Publisher
- American Physiological Society
- Published
- 2017-03-01
- Volume
- 312
- Issue
- 3
- Pages
- C316-C327
- ISSN
- 0363-6143 (ISSN-L)
- 1522-1563
- OpenAlex
- W2567361543
- Cited by
- 50
- References
- 43
- Citations by year
- 2026: 2 · 2025: 6 · 2024: 2 · 2023: 3 · 2022: 5 · 2021: 1 · 2020: 8 · 2019: 7 · 2018: 12 · 2017: 4
- FWCI
- 6.17
- Type
- journal article
- Language
- en
- Open access
- closed
- Locations
- American Journal of Physiology-Cell Physiology (publishedVersion)
- PubMed (publishedVersion)
- Victoria University Research Repository (Victoria University) (submittedVersion)
- Funders
- Department of Health, Australian Government | National Health and Medical Research Council (NHMRC) (10551460)
- Topics
- Health Sciences › Medicine › Physiology › Nitric Oxide and Endothelin Effects
- Health Sciences › Medicine › Cardiology and Cardiovascular Medicine › Cardiomyopathy and Myosin Studies
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Redox biology and oxidative stress
- Keywords
- S-nitrosylation, S-glutathionylation, troponin I, calcium sensitivity, fast-twitch muscle, nitric oxide, skeletal muscle contraction, S-nitroso-N-acetylpenicillamine, S-nitrosoglutathione, N-ethylmaleimide, type I muscle fibers, mechanomyogram
- MeSH terms
- Animals, Binding Sites, Calcium, Cells, Cultured, Chickens, Cysteine, Glutathione, Humans, Isometric Contraction, Male, Nitric Oxide, Protein Binding, Species Specificity, Rats, Sprague-Dawley, Muscle Fibers, Fast-Twitch, Troponin I, Calcium Signaling, Rats, Long-Evans, Rats, Young Adult
About these checks
Registry, retraction, Cites and statistics results are computed, not judged, with one exception: a printed reference that no index lists is read once by a model before it counts as a citation error. Claims are extracted by a model and kept only when their quote appears verbatim in the text; each cited source is then checked, with a model’s help, for whether it says what the claim relies on. Those verdicts can be wrong — read the claim’s quote and the cited source before relying on one.
Statistics and arithmetic are recomputed, not judged.
Confidence bands are shown only for fields calibrated on our annotated test set. None is calibrated yet, so no band is shown.
What counts as a claim ›How we measure confidence ›How we check this ›
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