Abstract
The coupling of hemoglobin sensing of physiological oxygen gradients to stimulation of nitric oxide (NO) bioactivity is an established principle of hypoxic blood flow. One mechanism proposed to explain this oxygen-sensing–NO bioactivity linkage postulates an essential role for the conserved Cys93 residue of the hemoglobin β-chain (βCys93) and, specifically, for S-nitrosation of βCys93 to form S-nitrosohemoglobin (SNO-Hb)1. The SNO-Hb hypothesis, which conceptually links hemoglobin and NO biology, has been debated intensely in recent years2,3. This debate has precluded a consensus on physiological mechanisms and on assessment of the potential role of SNO-Hb in pathology. Here we describe new mouse models that exclusively express either human wild-type hemoglobin or human hemoglobin in which the βCys93 residue is replaced with alanine to assess the role of SNO-Hb in red blood cell–mediated hypoxic vasodilation. Substitution of this residue, precluding hemoglobin S-nitrosation, did not change total red blood cell S-nitrosothiol abundance but did shift S-nitrosothiol distribution to lower molecular weight species, consistent with the loss of SNO-Hb. Loss of βCys93 resulted in no deficits in systemic or pulmonary hemodynamics under basal conditions and, notably, did not affect isolated red blood cell–dependent hypoxic vasodilation. These results demonstrate that SNO-Hb is not essential for the physiologic coupling of erythrocyte deoxygenation with increased NO bioactivity in vivo.
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Acknowledgements
This study was supported by grants from the US National Institutes of Health (HL057619) to T.M.T., from the American Heart Association, Southeast Affiliate (AHA 0655312B) to R.P.P. and from the Alabama Neuroscience Blueprint Core Center (NS 057098). T.S.I. was supported by a Cardiovascular Pathophysiology Training Fellowship. We also thank T. Lowder and M. Hewitt for technical assistance in exercise studies.
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T.S.I., C.-W.S., L.-C.W., X.T., D.A.V. and K.M.P. were responsible for performing experiments. T.S.I., C.-W.S., D.A.V., R.P.P. and T.M.T. were responsible for planning all experiments, analyzing data and writing the manuscript. M.B.R. contributed to mass spectrometry assays, L.S. was responsible for exercise-related studies, C.G.K. and B.G.B. were responsible for capillary density measurements, N.P. and J.M.W. contributed to blood pressure measurements and N.A. assessed pulmonary hemodynamics. J.R. did the embryonic stem cell injections to generate the chimeras.
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Supplementary Table 1, Supplementary Figs. 1–7 and Supplementary Methods (PDF 1201 kb)
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Isbell, T., Sun, CW., Wu, LC. et al. SNO-hemoglobin is not essential for red blood cell–dependent hypoxic vasodilation. Nat Med 14, 773–777 (2008). https://doi.org/10.1038/nm1771
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DOI: https://doi.org/10.1038/nm1771
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