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Cited by 403Redox biology and oxidative stressNitric Oxide and Endothelin EffectsHemoglobin structure and function

Diffusion of peroxynitrite across erythrocyte membranes

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Denicola, Ana, Souza, José M., Radi, Rafael·1998·Proceedings of the National Academy of Sciences
DOI 10.1073/pnas.95.7.3566PMC PMC19876
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Abstract

Peroxynitrite anion (ONOO − ) is a reactive species of increasingly recognized biological relevance that contributes to oxidative tissue damage. At present, however, there is limited knowledge about the mechanisms of peroxynitrite diffusion through biological compartments. In this work we have studied the diffusion of peroxynitrite across erythrocyte membranes. In solution, peroxynitrite rapidly reacts with oxyhemoglobin to yield methemoglobin, with k 2 = (10.4 ± 0.3) × 10 3 M −1 ⋅s −1 at pH 7.4 and 25°C. Addition of peroxynitrite to intact erythrocytes caused oxidation of intracellular oxyhemoglobin to methemoglobin. Oxidation yields in red blood cells at pH 7.0 were approximately 40% of those obtained in solution, which results mostly from competition of other cytosolic components for peroxynitrite. Indeed, rather small differences were observed between oxidation yields in lysates compared with intact erythrocytes, in particular at acidic and neutral pH values, indicating that membrane was not precluding peroxynitrite diffusion. Incubation of erythrocytes at pH 7.0 with 4,4′-diisothiocyanatostilbene-2,2′-disulfonic acid (DIDS), a specific inhibitor of anion exchange, resulted in up to 50% inhibition of oxyhemoglobin oxidation by peroxynitrite. More protection by DIDS was achieved at alkaline pH, while no effect was observed at pH 5.5, where 95% of peroxynitrite is in the acidic form, ONOOH (pK a = 6.8). In addition, peroxynitrite caused nitration of intracellular hemoglobin, in a process that was enhanced in thiol-depleted erythrocytes. Our results indicate that peroxynitrite is able to cross the erythrocyte membrane by two different mechanisms: in the anionic form through the DIDS-inhibitable anion channel, and in the protonated form by passive diffusion.

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  • Peroxynitrite can mediate oxidation/nitration distant from its site of formation.

    “Additionally, studies show peroxynitrite can traverse cell membranes by both diffusion and through ion channels, suggesting that peroxynitrite can mediate oxidation/nitration distant from its site of formation (221, 611).”

    From Sources of Vascular Nitric Oxide and Reactive Oxygen Species and Their Regulation (2019) · cites this paper as [221]

  • Peroxynitrite can traverse cell membranes by both diffusion and through ion channels.

    “Additionally, studies show peroxynitrite can traverse cell membranes by both diffusion and through ion channels, suggesting that peroxynitrite can mediate oxidation/nitration distant from its site of formation (221, 611).”

    From Sources of Vascular Nitric Oxide and Reactive Oxygen Species and Their Regulation (2019) · cites this paper as [221]

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Research works printed but not linked: 7

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Authors
Denicola, A., Souza, J. M., & Radi, R.
Affiliations
  • Universidad de la República de Uruguay (UY)
  • Department of Physical Biochemistry, Facultad de Ciencias, and Department of Biochemistry, Facultad de Medicina, Universidad de la República, Montevideo, Uruguay
ORCID
Venue
Proceedings of the National Academy of Sciences
Publisher
National Academy of Sciences
Published
1998-03-31
Volume
95
Issue
7
Pages
3566-3571
ISSN
  • 0027-8424 (ISSN-L)
  • 1091-6490
DOI
10.1073/pnas.95.7.3566
PMC
PMC19876
OpenAlex
W2018380711
Cited by
403
References
55
Citations by year
2026: 4 · 2025: 6 · 2024: 6 · 2023: 6 · 2022: 17 · 2021: 13 · 2020: 10 · 2019: 18 · 2018: 11 · 2017: 8 · 2016: 12 · 2015: 15 · 2014: 16 · 2013: 13 · 2012: 21
FWCI
6.28
Type
journal article
Language
en
Open access
Locations
  • Proceedings of the National Academy of Sciences (publishedVersion)
  • Europe PMC (PubMed Central) (publishedVersion)
  • PubMed Central (publishedVersion)
  • PubMed (publishedVersion)
Topics
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Redox biology and oxidative stress
  • Health Sciences › Medicine › Physiology › Nitric Oxide and Endothelin Effects
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Cell Biology › Hemoglobin structure and function
Keywords
peroxynitrite, erythrocyte membrane, membrane permeability, methemoglobin, DIDS, anion channels, passive diffusion, pH dependence, thiol depletion, reactive oxygen species, oxidative tissue damage
MeSH terms
Biological Transport, Erythrocyte Membrane, Humans, Hydrogen-Ion Concentration, Nitrates, Oxidation-Reduction, Oxidants, 4,4'-Diisothiocyanostilbene-2,2'-Disulfonic Acid

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