Abstract
The tenet of high-affinity nitric oxide (NO) binding to a haemoglobin (Hb) has shaped our view of haem proteins and of small diffusible signaling molecules. Specifically, NO binds rapidly to haem iron in Hb (k ≈ 107 M−1 s−1) (refs 1, 2) and once bound, the NO activity is largely irretrievable (Kd ≈ 10−5 s−1) (3–10); the binding is purportedly so tight as to be unaffected by O2 or CO. However, these general principles do not consider the allosteric state of Hb or the nature of the allosteric effector, and they mostly derive from the functional behaviour of fully nitrosylated Hb, whereas Hb is only partially nitrosylated in vivo11,12,13,14,15,16. Here we show that oxygen drives the conversion of nitrosylhaemoglobin in the ‘tense’ T (or partially nitrosylated, deoxy) structure to S -nitrosohaemoglobin in the ‘relaxed’ R (or ligand-bound, oxy) structure. In the absence of oxygen, nitroxyl anion (NO−) is liberated in a reaction producing methaemoglobin. The yields of both S -nitrosohaemoglobin and methaemoglobin are dependent on the NO/Hb ratio. These newly discovered reactions elucidate mechanisms underlying NO function in the respiratory cycle, and provide insight into the aetiology of S -nitrosothiols, methaemoglobin and its related valency hybrids. Mechanistic re-examination of NO interactions with other haem proteins containing allosteric-site thiols may be warranted.
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References
Gibson, Q. H. & Rougton, F. J. W. The kinetics and equilibria of the reactions of nitric oxide with sheep haemoglobin. J. Physiol. 136, 507–526 (1957).
Cassoly, R. & Gibson, Q. H. Conformation, co-operativity and ligand binding in human hemoglobin. J. Mol. Biol. 91, 301–313 (1975).
Sharma, V. S. & Ranney, H. M. The dissociation of NO from nitrosylhemoglobin. J. Biol. Chem. 253, 6467–6472 (1978).
Moore, E. G. & Gibson, Q. H. Cooperativity in the dissociation of nitric oxide from hemoglobin. J. Biol. Chem. 251, 2788–2794 (1976).
Kharitonov, V. G., Bonaventura, J. & Sharma, V. S. in Methods in Nitric Oxide Research (eds Feelisch, M. & Stamler, J. S.) 39–47 (Wiley, London, (1996)).
Traylor, T. G. & Sharma, V. S. Why NO? Biochemistry 31, 2847–2849 (1992).
Lancaster, J. R. Simulation of the diffusion and reaction of endogenously produced nitric oxide. Proc. Natl Acad. Sci. USA 91, 8137–8141 (1994).
Motterlini, R., Vandegriff, K. D. & Winslow, R. M. Hemoglobin–nitric oxide interaction and its implications. Transfus. Med. Rev. X, 77–84 (1996).
Marletta, M. A., Tayeh, M. A. & Hevel, J. M. Unraveling the biological significance of nitric oxide. Biofactors 2, 219–225 (1990).
Antonini, E. & Brunori, M. in Hemoglobin and Myoglobin in their Reactions with Ligands 32, 272–276 (Elsevier, New York, (1971)).
Jia, L., Bonaventura, C., Bonaventura, J. & Stamler, J. S. S -nitrosohemoglobin: a dynamic activity of blood involved in vascular control. Nature 380, 221–226 (1996).
Stamler, J. S. et al. Blood flow regulation by S -nitrosohemoglobin is controlled by the physiological oxygen gradient. Science 276, 2034–2037 (1997).
Westenberger, U. et al. Formation of free radicals and nitric oxide derivative of hemoglobin in rats during septic shock. Free Rad. Res. Commun. 11, 167–178 (1990).
Hall, D. M., Buettner, G. R., Mathes, R. D. & Gisolfi, C. V. Hyperthermia stimulates nitric oxide formation: electron paramagnetic resonance detection of NO-heme in blood. J. Appl. Physiol. 77, 548–553 (1994).
Kruszyna, R., Kruszyna, H., Smith, R. P. & Wilcox, D. E. Generation of valency hybrids and nitrosylated species of hemoglobin in mice by nitric oxide vasodilators. Toxicol. Appl. Pharmacol. 94, 458–465 (1988).
Kosaka, H. et al. ESR spectral transition by arteriovenous cycle in nitric oxide hemoglobin of cytokine-treated rats. Am. J. Physiol. 266, 1400–1405 (1994).
Perutz, M. F. in Molecular Basis of Blood Diseases (ed. Stammatayanopoulos, G.) 127–178 (Saunders, Philadelphia, (1987)).
Taketa, F., Antholine, W. E. & Chen, J. Y. Chain nonequivalence in binding of nitric oxide to hemoglobin. J. Biol. Chem. 253, 5448–5451 (1978).
Henry, Y. et al. EPR characterization of molecular targets for NO in mammalian cells and organelles. FASEB J. 7, 1124–1134 (1993).
Addison, A. W. & Stephanos, J. J. Nitrosyliron (III) hemoglobin: autoreduction and spectroscopy. Biochemistry 25, 4104–4113 (1986).
Stamler, J. S., Singel, D. J., Loscalzo, J. Biochemistry of nitric oxide and its redox activated forms. Science 258, 1898–1902 (1992).
Arnelle, D. & Stamler, J. S. NO+, NO·, and NO− donation by S -nitrosothiols: Implications for regulation of physiological functions by S -nitrosylation and acceleration of disulfide formation. Archiv. Biochem. Biophys. 318, 279–285 (1995).
Kruszyna, R., Kruszyna, H., Smith, R. P., Thron, R. D. & Wilcox, D. E. Nitrite conversion to nitric oxide in red cells and its stabilization as a nitrosylated valency hybrid of hemoglobin. J. Pharmacol. Exp. Ther. 241, 307–313 (1987).
Kagan, V. E., Day, B. W., Elsayed, N. V. & Gorbunov, N. V. Dynamics of nitrosylated haemoglobin in blood. Nature 383, 30–31 (1996).
Riggs, A. & Wolbach, R. A. Sulfhydryl groups and the structure of hemoglobin. J. Gen. Physiol. 39, 585–605 (1956).
Riggs, A. F. The binding of N -ethylmaleimide by human hemoglobin and its effect upon the oxygen equilibrium. J. Biol. Chem. 236, 1948–1954 (1961).
Gow, A., Buerk, D. D. & Ischiropolouus, H. Anovel reaction mechanism for the formation of S -nitrosothiol in vivo. J. Biol. Chem. 272, 2841–2845 (1997).
Perutz, M. F. Blood: Taking the pressure off. Nature 380, 205–206 (1996).
Stamler, J. S., Toone, E. J., Lipton, S. A. & Sucher, N. J. (S)NO Signals: Translocation, regulation, and a consensus motif. Neuron 18, 691–696 (1997).
Stamler, J. S. Redox signalling: Nitrosylation and related target interactions of nitric oxide. Cell 78, 931–936 (1994).
Acknowledgements
We thank J. Bonaventura, D. J. Singel, H. Ishiropolous and I. Fridovich for discussion and T. McMahon for help with measurements. J.S.S. is the recipient of grants from the NHLBI; A.J.G. is supported by a National Research Award from the NHLBI.
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Gow, A., Stamler, J. Reactions between nitric oxide and haemoglobin under physiological conditions. Nature 391, 169–173 (1998). https://doi.org/10.1038/34402
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DOI: https://doi.org/10.1038/34402
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