Mechanisms of Human Erythrocytic Bioactivation of Nitrite
Open accessAbstract
Nitrite signaling likely occurs through its reduction to nitric oxide (NO). Several reports support a role of erythrocytes and hemoglobin in nitrite reduction, but this remains controversial, and alternative reductive pathways have been proposed. In this work we determined whether the primary human erythrocytic nitrite reductase is hemoglobin as opposed to other erythrocytic proteins that have been suggested to be the major source of nitrite reduction. We employed several different assays to determine NO production from nitrite in erythrocytes including electron paramagnetic resonance detection of nitrosyl hemoglobin, chemiluminescent detection of NO, and inhibition of platelet activation and aggregation. Our studies show that NO is formed by red blood cells and inhibits platelet activation. Nitric oxide formation and signaling can be recapitulated with isolated deoxyhemoglobin. Importantly, there is limited NO production from erythrocytic xanthine oxidoreductase and nitric-oxide synthase. Under certain conditions we find dorzolamide (an inhibitor of carbonic anhydrase) results in diminished nitrite bioactivation, but the role of carbonic anhydrase is abrogated when physiological concentrations of CO2 are present. Importantly, carbon monoxide, which inhibits hemoglobin function as a nitrite reductase, abolishes nitrite bioactivation. Overall our data suggest that deoxyhemoglobin is the primary erythrocytic nitrite reductase operating under physiological conditions and accounts for nitrite-mediated NO signaling in blood.
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The ratio of NADPH to l-citrulline is close to the optimal value of 1.5 molecules of NADPH per l-citrulline (and NO) generated.
“The electrons transferred from NADPH are almost quantitatively used to catalyze l-arginine oxidation, and the ratio of NADPH to l-citrulline is close to the optimal value of 1.5 molecules of NADPH per l-citrulline (and NO) generated (3, 590).”
From Sources of Vascular Nitric Oxide and Reactive Oxygen Species and Their Regulation (2019) · cites this paper as [590]
Deoxyhemoglobin may be the only nitrite reductase of physiological relevance under hypoxic conditions, as inactivation of deoxyhemoglobin by treatment with carbon monoxide completely blocked NO release from RBCs as assessed by electron paramagnetic resonance.
“However, it was shown recently that deoxyhemoglobin may be the only nitrite reductase of physiological relevance under hypoxic conditions (Eq. 17) as inactivation of deoxyhemoglobin by treatment with carbon monoxide completely blocked NO release from RBCs as assessed by electron paramagnetic resonance (107).”
From Red Blood Cell Function and Dysfunction: Redox Regulation, Nitric Oxide Metabolism, Anemia (2017) · cites this paper as [107]
Deoxyhemoglobin is the primary erythrocytic nitrite reductase under physiological conditions.
“The presumption that deoxyhemoglobin was the primary erythrocytic nitrite reductase under physiological conditions was recently confirmed [79].”
From Signaling and stress: The redox landscape in NOS2 biology (2015) · cites this paper as [79]
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- Authors
- Liu, C., Wajih, N., Liu, X., Basu, S., Janes, J., Marvel, M., Keggi, C., Helms, C. C., Lee, A. N., Belanger, A. M., Diz, D. I., Laurienti, P. J., Caudell, D. L., Wang, J., Gladwin, M. T., & Kim-Shapiro, D. B.
- Affiliations
- Wake Forest University Health Sciences (US)
- University of Richmond (US)
- Wake Forest University (US)
- University of Pittsburgh (US)
- ORCID
- Venue
- Journal of Biological Chemistry
- Publisher
- Elsevier BV
- Published
- 2015-01
- Volume
- 290
- Issue
- 2
- Pages
- 1281-1294
- ISSN
- 0021-9258 (ISSN-L)
- PMC
- PMC4294492
- OpenAlex
- W2012574454
- Cited by
- 76
- References
- 20
- Citations by year
- 2026: 1 · 2025: 2 · 2024: 2 · 2023: 3 · 2022: 4 · 2021: 5 · 2020: 6 · 2019: 5 · 2018: 10 · 2017: 10 · 2016: 19 · 2015: 8 · 2014: 1
- FWCI
- 6.31
- Type
- journal article
- Language
- en
- Open access
- hybrid
- doi.org
- Licence
- Locations
- Journal of Biological Chemistry (publishedVersion, cc-by)
- Europe PMC (PubMed Central) (publishedVersion)
- PubMed Central (publishedVersion)
- PubMed (publishedVersion)
- Funders
- National Institutes of Health (T32 GM095440, HL058091, HL098032, P30 CA121291-37)
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Cell Biology › Hemoglobin structure and function
- Health Sciences › Medicine › Physiology › Nitric Oxide and Endothelin Effects
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Heme Oxygenase-1 and Carbon Monoxide
- Keywords
- nitric oxide production, erythrocytes, hemoglobin, deoxyhemoglobin, nitrite reductase, platelet activation, platelet aggregation, xanthine oxidoreductase, nitric oxide synthase, carbonic anhydrase, carbon monoxide, carbon dioxide
- MeSH terms
- Blood Platelets, Carbonic Anhydrases, Electron Spin Resonance Spectroscopy, Erythrocytes, Hemoglobins, Humans, Nitric Oxide, Nitrite Reductases, Nitrites, Oxidation-Reduction, Sulfonamides, Thiophenes, Signal Transduction
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