Intermediates Detected by Visible Spectroscopy during the Reaction of Nitrite with Deoxyhemoglobin: The Effect of Nitrite Concentration and Diphosphoglycerate
Abstract
The reaction of nitrite with deoxyhemoglobin (deoxyHb) results in the reduction of nitrite to NO, which binds unreacted deoxyHb forming Fe(II)-nitrosylhemoglobin (Hb(II)NO). The tight binding of NO to deoxyHb is, however, inconsistent with reports implicating this reaction with hypoxic vasodilation. This dilemma is resolved by the demonstration that metastable intermediates are formed in the course of the reaction of nitrite with deoxyHb. The level of intermediates is quantitated by the excess deoxyHb consumed over the concentrations of the final products formed. The dominant intermediate has a spectrum that does not correspond to that of Hb(III)NO formed when NO reacts with methemoglobin (MetHb), but is similar to metHb resulting in the spectroscopic determinations of elevated levels of metHb. It is a delocalized species involving the heme iron, the NO, and perhaps the beta-93 thiol. The putative role for red cell reacted nitrite on vasodilation is associated with reactions involving the intermediate. (1) The intermediate is less stable with a 10-fold excess of nitrite and is not detected with a 100-fold excess of nitrite. This observation is attributed to the reaction of nitrite with the intermediate producing N2O3. (2) The release of NO quantitated by the formation of Hb(II)NO is regulated by changes in the distal heme pocket as shown by the 4.5-fold decrease in the rate constant in the presence of 2,3-diphosphoglycerate. The regulated release of NO or N2O3 as well as the formation of the S-nitroso derivative of hemoglobin, which has also been reported to be formed from the intermediates generated during nitrite reduction, should be associated with any hypoxic vasodilation attributed to the RBC.
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Through these signaling mechanisms, nitrated lipids mediate a number of physiological effects in animal models including attenuation of inflammatory bowel disease, protection after ischemia/reperfusion, decreases in atherosclerosis, and reversal of obesity-induced pulmonary hypertension.
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- Authors
- Nagababu, E., Ramasamy, S., & Rifkind, J. M.
- Affiliations
- National Institutes of Health (US)
- Corresponding author
- Rifkind, Joseph M.
- Venue
- Biochemistry
- Publisher
- American Chemical Society (ACS)
- Published
- 2007-09-20
- Volume
- 46
- Issue
- 41
- Pages
- 11650-11659
- ISSN
- 0006-2960 (ISSN-L)
- 1520-4995
- OpenAlex
- W2030738199
- Cited by
- 33
- References
- 10
- Citations by year
- 2025: 1 · 2023: 1 · 2019: 2 · 2018: 1 · 2016: 3 · 2014: 3 · 2013: 3 · 2012: 4
- FWCI
- 1.38
- Type
- journal article
- Language
- en
- Open access
- closed
- Locations
- Biochemistry (publishedVersion)
- PubMed (publishedVersion)
- Funders
- National Institutes of Health
- 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
- nitrite, deoxyhemoglobin, nitric oxide, metastable intermediates, methemoglobin, N2O3, hypoxic vasodilation, erythrocytes, S-nitrosohemoglobin, distal heme pocket, nitrite reduction
- MeSH terms
- Diphosphoglyceric Acids, Hemoglobins, Humans, Kinetics, Nitric Oxide, Nitrites, Spectrophotometry, Luminescence
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