Relative Role of Heme Nitrosylation and Beta-cysteine 93 Nitrosation in the Transport and Metabolism of Nitric Oxide by Hemoglobin in the Human Circulation

Overview

Journal Proc Natl Acad Sci U S A

Specialty Science

Date 2000 Aug 24

PMID 10954746

Citations 63

Authors

M T Gladwin

F P Ognibene

L K Pannell

J S Nichols

J H Shelhamer

A N Schechter

Affiliations

Soon will be listed here.

Abstract

To quantify the reactions of nitric oxide (NO) with hemoglobin under physiological conditions and to test models of NO transport on hemoglobin, we have developed an assay to measure NO-hemoglobin reaction products in normal volunteers, under basal conditions and during NO inhalation. NO inhalation markedly raised total nitrosylated hemoglobin levels, with a significant arterial-venous gradient, supporting a role for hemoglobin in the transport and delivery of NO. The predominant species accounting for this arterial-venous gradient is nitrosyl(heme)hemoglobin. NO breathing increases S-nitrosation of hemoglobin beta-chain cysteine 93, however only to a fraction of the level of nitrosyl(heme)hemoglobin and without a detectable arterial-venous gradient. A strong correlation between methemoglobin and plasma nitrate formation was observed, suggesting that NO metabolism is a primary physiological cause of hemoglobin oxidation. Our results demonstrate that NO-heme reaction pathways predominate in vivo, NO binding to heme groups is a rapidly reversible process, and S-nitrosohemoglobin formation is probably not a primary transport mechanism for NO but may facilitate NO release from heme.

Citing Articles

Hemoglobin Variants as Targets for Stabilizing Drugs.

Zoldakova M, Novotny M, Khakurel K, Zoldak G Molecules. 2025; 30(2).

PMID: 39860253 PMC: 11767434. DOI: 10.3390/molecules30020385.

The Neural Palette of Heme: Altered Heme Homeostasis Underlies Defective Neurotransmission, Increased Oxidative Stress, and Disease Pathogenesis.

Soladogun A, Zhang L Antioxidants (Basel). 2025; 13(12.

PMID: 39765770 PMC: 11672823. DOI: 10.3390/antiox13121441.

Nitric oxide and thiols: Chemical biology, signalling paradigms and vascular therapeutic potential.

Dent M, DeMartino A Br J Pharmacol. 2023; .

PMID: 37908126 PMC: 11058123. DOI: 10.1111/bph.16274.

How Nitric Oxide Hindered the Search for Hemoglobin-Based Oxygen Carriers as Human Blood Substitutes.

Samaja M, Malavalli A, Vandegriff K Int J Mol Sci. 2023; 24(19).

PMID: 37834350 PMC: 10573492. DOI: 10.3390/ijms241914902.

A Molecular Troika of Angiogenesis, Coagulopathy and Endothelial Dysfunction in the Pathology of Avascular Necrosis of Femoral Head: A Comprehensive Review.

Singh M, Singh B, Sharma K, Kumar N, Mastana S, Singh P Cells. 2023; 12(18).

PMID: 37759498 PMC: 10528276. DOI: 10.3390/cells12182278.

References

Bonaventura C, Godette G, Tesh S, Holm D, Bonaventura J, Crumbliss A . Internal electron transfer between hemes and Cu(II) bound at cysteine beta93 promotes methemoglobin reduction by carbon monoxide. J Biol Chem. 1999; 274(9):5499-507. DOI: 10.1074/jbc.274.9.5499. View

Fang K, Ragsdale N, Carey R, MacDonald T, Gaston B . Reductive assays for S-nitrosothiols: implications for measurements in biological systems. Biochem Biophys Res Commun. 1998; 252(3):535-40. DOI: 10.1006/bbrc.1998.9688. View

Cooper C . Nitric oxide and iron proteins. Biochim Biophys Acta. 1999; 1411(2-3):290-309. DOI: 10.1016/s0005-2728(99)00021-3. View

Kosaka H . Nitric oxide and hemoglobin interactions in the vasculature. Biochim Biophys Acta. 1999; 1411(2-3):370-7. DOI: 10.1016/s0005-2728(99)00026-2. View

Patel R, Hogg N, Spencer N, Kalyanaraman B, Matalon S, Darley-Usmar V . Biochemical characterization of human S-nitrosohemoglobin. Effects on oxygen binding and transnitrosation. J Biol Chem. 1999; 274(22):15487-92. DOI: 10.1074/jbc.274.22.15487. View

Gow A, Luchsinger B, Pawloski J, Singel D, Stamler J . The oxyhemoglobin reaction of nitric oxide. Proc Natl Acad Sci U S A. 1999; 96(16):9027-32. PMC: 17726. DOI: 10.1073/pnas.96.16.9027. View

Kubes P, Payne D, Grisham M, Fox-Robichaud A . Inhaled NO impacts vascular but not extravascular compartments in postischemic peripheral organs. Am J Physiol. 1999; 277(2):H676-82. DOI: 10.1152/ajpheart.1999.277.2.H676. View

Bonaventura C, Ferruzzi G, Tesh S, Stevens R . Effects of S-nitrosation on oxygen binding by normal and sickle cell hemoglobin. J Biol Chem. 1999; 274(35):24742-8. DOI: 10.1074/jbc.274.35.24742. View

Gross S, Lane P . Physiological reactions of nitric oxide and hemoglobin: a radical rethink. Proc Natl Acad Sci U S A. 1999; 96(18):9967-9. PMC: 33713. DOI: 10.1073/pnas.96.18.9967. View

10.

Stubauer G, Giuffre A, Sarti P . Mechanism of S-nitrosothiol formation and degradation mediated by copper ions. J Biol Chem. 1999; 274(40):28128-33. DOI: 10.1074/jbc.274.40.28128. View

11.

Wolzt M, MacAllister R, Davis D, Feelisch M, Moncada S, Vallance P . Biochemical characterization of S-nitrosohemoglobin. Mechanisms underlying synthesis, no release, and biological activity. J Biol Chem. 1999; 274(41):28983-90. DOI: 10.1074/jbc.274.41.28983. View

12.

Gladwin M, Schechter A, Shelhamer J, Pannell L, Conway D, Hrinczenko B . Inhaled nitric oxide augments nitric oxide transport on sickle cell hemoglobin without affecting oxygen affinity. J Clin Invest. 1999; 104(7):937-45. PMC: 408559. DOI: 10.1172/JCI7637. View

13.

Schoenborn B . Binding of xenon to horse haemoglobin. Nature. 1965; 208(5012):760-2. DOI: 10.1038/208760a0. View

14.

Winterbourn C, Carrell R . Oxidation of human haemoglobin by copper. Mechanism and suggested role of the thiol group of residue beta-93. Biochem J. 1977; 165(1):141-8. PMC: 1164879. DOI: 10.1042/bj1650141. View

15.

Huang T . NMR studies of the quaternary structure and heterogeneity of nitrosyl- and methemoglobin. J Biol Chem. 1979; 254(22):11467-74. View

16.

Wade R, Castro C . Redox reactivity of iron(III) porphyrins and heme proteins with nitric oxide. Nitrosyl transfer to carbon, oxygen, nitrogen, and sulfur. Chem Res Toxicol. 1990; 3(4):289-91. DOI: 10.1021/tx00016a002. View

17.

Wink D, Darbyshire J, Nims R, Saavedra J, Ford P . Reactions of the bioregulatory agent nitric oxide in oxygenated aqueous media: determination of the kinetics for oxidation and nitrosation by intermediates generated in the NO/O2 reaction. Chem Res Toxicol. 1993; 6(1):23-7. DOI: 10.1021/tx00031a003. View

18.

Kosaka H, Sawai Y, Sakaguchi H, Kumura E, Harada N, Watanabe M . ESR spectral transition by arteriovenous cycle in nitric oxide hemoglobin of cytokine-treated rats. Am J Physiol. 1994; 266(5 Pt 1):C1400-5. DOI: 10.1152/ajpcell.1994.266.5.C1400. View

19.

Wink D, Nims R, Darbyshire J, Christodoulou D, Hanbauer I, Cox G . Reaction kinetics for nitrosation of cysteine and glutathione in aerobic nitric oxide solutions at neutral pH. Insights into the fate and physiological effects of intermediates generated in the NO/O2 reaction. Chem Res Toxicol. 1994; 7(4):519-25. DOI: 10.1021/tx00040a007. View

20.

Zweier J, Wang P, Samouilov A, Kuppusamy P . Enzyme-independent formation of nitric oxide in biological tissues. Nat Med. 1995; 1(8):804-9. DOI: 10.1038/nm0895-804. View