Cited by 160Hemoglobin structure and functionNitric Oxide and Endothelin EffectsHeme Oxygenase-1 and Carbon Monoxide

S-Nitrosohemoglobin Is Unstable in the Reductive Erythrocyte Environment and Lacks O2/NO-linked Allosteric Function

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Gladwin, Mark T., Wang, Xunde, Reiter, Christopher D., Yang, Benjamin K., Vivas, Esther X., Bonaventura, Celia, Schechter, Alan N.·2002·Journal of Biological Chemistry

Abstract

Our previous results run counter to the hypothesis that S-nitrosohemoglobin (SNO-Hb) serves as an in vivo reservoir for NO from which NO release is allosterically linked to oxygen release. We show here that SNO-Hb undergoes reductive decomposition in erythrocytes, whereas it is stable in purified solutions and in erythrocyte lysates treated with an oxidant such as ferricyanide. Using an extensively validated methodology that eliminates background nitrite and stabilizes erythrocyte S-nitrosothiols, we find the levels of SNO-Hb in the basal human circulation, including red cell membrane fractions, were 46 +/- 17 nm in human arterial erythrocytes and 69 +/- 11 nm in venous erythrocytes, incompatible with the postulated reservoir function of SNO-Hb. Moreover, we performed experiments on human red blood cells in which we elevated the levels of SNO-Hb to 10,000 times the normal in vivo levels. The elevated levels of intra-erythrocytic SNO-Hb fell rapidly, independent of oxygen tension and hemoglobin saturation. Most of the NO released during this process was oxidized to nitrate. A fraction (25%) was exported as S-nitrosothiol, but this fraction was not increased at low oxygen tensions that favor the deoxy (T-state) conformation of Hb. Results of these studies show that, within the redox-active erythrocyte environment, the beta-globin cysteine 93 is maintained in a reduced state, necessary for normal oxygen affinity, and incapable of oxygen-linked NO storage and delivery.

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  • NO bioactivity in blood and tissue is preserved in the form of nitrite, nitrate, S-nitrosothiols, N-nitrosamines, nitrosylhemoglobin, and S-nitrosohemoglobin.

    “However studies conducted in this area over the last few years have unraveled mechanisms by which NO bioactivity in blood and in tissue have been preserved in the form of nitrite, nitrate, S-nitrosothiols (RSNO), N-nitrosamines (RNNO), nitrosylhemoglobin (NOHb), and S-nitrosohemoglobin (SNOHb) [16–21].”

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Research works printed but not linked: 7

  1. Ignarro, L. J., Buga, G. M., Wood, K. S., Byrns, R. E., and Chaudhuri, G. (1987) Proc. Natl. Acad. Sci. U. S. A. 84, 9265-92659
  2. Gladwin, M. T., Shelhamer, J. H., Schechter, A. N., Pease-Fye, M. E., Waclawiw, M. A., Panza, J. A., Ognibene, F. P., and Cannon, R. O., III (2000) Proc. Natl. Acad. Sci. U. S. A. 97, 11482-11487
  3. Stamler, J. S., Jaraki, O., Osborne, J., Simon, D. I., Keaney, J., Vita, J., Singel, D., Valeri, C. R., and Loscalzo, J. (1992) Proc. Natl. Acad. Sci. U. S. A. 89, 7674 -7677
  4. Gladwin, M. T., Ognibene, F. P., Pannell, L. K., Nichols, J. S., Pease-Fye, M. E., Shelhamer, J. H., and Schechter, A. N. (2000) Proc. Natl. Acad. Sci.
  5. Gaston, B., Reilly, J., Drazen, J. M., Fackler, J., Ramdev, P., Arnelle, D., Mullins, M. E., Sugarbaker, D. J., Chee, C., Singel, D. J., Loscalzo, J., and Stamler, J. S. (1993) Proc. Natl. Acad. Sci. U. S. A. 90, 10957-10961
  6. Nedospasov, A., Rafikov, R., Beda, N., and Nudler, E. (2000) Proc. Natl. Acad. Sci. U. S. A. 97, 13543-13548
  7. Huang, K. T., Han, T. H., Hyduke, D. R., Vaughn, M. W., Van Herle, H., Hein, T. W., Zhang, C., Kuo, L., and Liao, J. C. (2001) Proc. Natl. Acad. Sci.

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Authors
Gladwin, M. T., Wang, X., Reiter, C. D., Yang, B. K., Vivas, E. X., Bonaventura, C., & Schechter, A. N.
Affiliations
  • National Institutes of Health (US)
  • Duke University (US)
Corresponding author
Gladwin, Mark T.
Venue
Journal of Biological Chemistry
Publisher
Elsevier BV
Published
2002-08
Volume
277
Issue
31
Pages
27818-27828
ISSN
0021-9258 (ISSN-L)
OpenAlex
W2061156310
Cited by
160
References
45
Citations by year
2026: 1 · 2024: 2 · 2023: 3 · 2022: 3 · 2021: 1 · 2020: 1 · 2019: 1 · 2018: 4 · 2017: 4 · 2016: 3 · 2015: 1 · 2014: 3 · 2013: 4 · 2012: 5
FWCI
7.92
Type
journal article
Language
en
Open access
Locations
  • Journal of Biological Chemistry (publishedVersion, cc-by)
  • PubMed (publishedVersion)
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
S-nitrosohemoglobin, nitric oxide storage, erythrocytes, S-nitrosothiols, reductive decomposition, oxygen affinity, allosteric regulation, nitrate formation, redox conditions, nitric oxide delivery
MeSH terms
Allosteric Regulation, Animals, Cattle, Drug Stability, Erythrocytes, Hemoglobins, Humans, Kinetics, Nitric Oxide, Nitrites, Oxygen, Serum Albumin, Serum Albumin, Bovine

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