Endothelial cell expression of haemoglobin α regulates nitric oxide signalling
Adam C Straub, Alexander W Lohman, Marie Billaud, Scott R Johnstone, Scott T Dwyer, Monica Y Lee, Pamela Schoppee Bortz, Angela K Best, Linda Columbus, Benjamin Gaston, Brant E Isakson, Adam C Straub, Alexander W Lohman, Marie Billaud, Scott R Johnstone, Scott T Dwyer, Monica Y Lee, Pamela Schoppee Bortz, Angela K Best, Linda Columbus, Benjamin Gaston, Brant E Isakson
Abstract
Models of unregulated nitric oxide (NO) diffusion do not consistently account for the biochemistry of NO synthase (NOS)-dependent signalling in many cell systems. For example, endothelial NOS controls blood pressure, blood flow and oxygen delivery through its effect on vascular smooth muscle tone, but the regulation of these processes is not adequately explained by simple NO diffusion from endothelium to smooth muscle. Here we report a new model for the regulation of NO signalling by demonstrating that haemoglobin (Hb) α (encoded by the HBA1 and HBA2 genes in humans) is expressed in human and mouse arterial endothelial cells and enriched at the myoendothelial junction, where it regulates the effects of NO on vascular reactivity. Notably, this function is unique to Hb α and is abrogated by its genetic depletion. Mechanistically, endothelial Hb α haem iron in the Fe(3+) state permits NO signalling, and this signalling is shut off when Hb α is reduced to the Fe(2+) state by endothelial cytochrome b5 reductase 3 (CYB5R3, also known as diaphorase 1). Genetic and pharmacological inhibition of CYB5R3 increases NO bioactivity in small arteries. These data reveal a new mechanism by which the regulation of the intracellular Hb α oxidation state controls NOS signalling in non-erythroid cells. This model may be relevant to haem-containing globins in a broad range of NOS-containing somatic cells.
Conflict of interest statement
Competing financial interest
None
Figures
References
- Lim KH, Ancrile BB, Kashatus DF, Counter CM. Tumour maintenance is mediated by eNOS. Nature. 2008;452:646–649. doi: 10.1038/nature06778. nature06778 [pii]
- Hess DT, Matsumoto A, Kim SO, Marshall HE, Stamler JS. Protein S-nitrosylation: purview and parameters. Nat Rev Mol Cell Biol. 2005;6:150–166. doi: 10.1038/nrm1569. nrm1569 [pii]
- Bolotina VM, Najibi S, Palacino JJ, Pagano PJ, Cohen RA. Nitric oxide directly activates calcium-dependent potassium channels in vascular smooth muscle. Nature. 1994;368:850–853. doi: 10.1038/368850a0.
- Shesely EG, et al. Elevated blood pressures in mice lacking endothelial nitric oxide synthase. Proc Natl Acad Sci U S A. 1996;93:13176–13181.
- Straub AC, et al. Compartmentalized connexin 43 s-nitrosylation/denitrosylation regulates heterocellular communication in the vessel wall. Arterioscler Thromb Vasc Biol. 2011;31:399–407. doi: 10.1161/ATVBAHA.110.215939. ATVBAHA.110.215939 [pii]
- Hultquist DE, Passon PG. Catalysis of methaemoglobin reduction by erythrocyte cytochrome B5 and cytochrome B5 reductase. Nat New Biol. 1971;229:252–254.
- Newton DA, Rao KM, Dluhy RA, Baatz JE. Hemoglobin is expressed by alveolar epithelial cells. J Biol Chem. 2006;281:5668–5676. doi: 10.1074/jbc.M509314200. M509314200 [pii]
- Nishi H, et al. Hemoglobin is expressed by mesangial cells and reduces oxidant stress. J Am Soc Nephrol. 2008;19:1500–1508. doi: 10.1681/ASN.2007101085. ASN.2007101085 [pii]
- Liu L, Zeng M, Stamler JS. Hemoglobin induction in mouse macrophages. Proc Natl Acad Sci U S A. 1999;96:6643–6647.
- Schelshorn DW, et al. Expression of hemoglobin in rodent neurons. J Cereb Blood Flow Metab. 2009;29:585–595. doi: 10.1038/jcbfm.2008.152. jcbfm2008152 [pii]
- Halligan KE, Jourd’heuil FL, Jourd’heuil D. Cytoglobin is expressed in the vasculature and regulates cell respiration and proliferation via nitric oxide dioxygenation. J Biol Chem. 2009;284:8539–8547. doi: 10.1074/jbc.M808231200. M808231200 [pii]
- Brunori M, et al. Neuroglobin, nitric oxide, and oxygen: functional pathways and conformational changes. Proc Natl Acad Sci U S A. 2005;102:8483–8488. doi: 10.1073/pnas.0408766102. 0408766102 [pii]
- Flogel U, Merx MW, Godecke A, Decking UK, Schrader J. Myoglobin: A scavenger of bioactive NO. Proc Natl Acad Sci U S A. 2001;98:735–740. doi: 10.1073/pnas.011460298011460298[pii].
- Dora KA, Doyle MP, Duling BR. Elevation of intracellular calcium in smooth muscle causes endothelial cell generation of NO in arterioles. Proceedings of the National Academy of Sciences of the United States of America. 1997;94:6529–6534.
- Angelo M, Hausladen A, Singel DJ, Stamler JS. Interactions of NO with hemoglobin: from microbes to man. Methods Enzymol. 2008;436:131–168. doi: 10.1016/S0076-6879(08)36008-X. S0076-6879(08)36008-X [pii]
- Gladwin MT, Lancaster JR, Jr, Freeman BA, Schechter AN. Nitric oxide’s reactions with hemoglobin: a view through the SNO-storm. Nat Med. 2003;9:496–500. doi: 10.1038/nm0503-496nm0503-496[pii].
- Palmer RM, Ferrige AG, Moncada S. Nitric oxide release accounts for the biological activity of endothelium-derived relaxing factor. Nature. 1987;327:524–526. doi: 10.1038/327524a0.
- Ignarro LJ, Adams JB, Horwitz PM, Wood KS. Activation of soluble guanylate cyclase by NO-hemoproteins involves NO-heme exchange. Comparison of heme-containing and heme-deficient enzyme forms. J Biol Chem. 1986;261:4997–5002.
- Ignarro LJ, Byrns RE, Buga GM, Wood KS. Endothelium-derived relaxing factor from pulmonary artery and vein possesses pharmacologic and chemical properties identical to those of nitric oxide radical. Circ Res. 1987;61:866–879.
- Cassoly R, Gibson Q. Conformation, co-operativity and ligand binding in human hemoglobin. J Mol Biol. 1975;91:301–313.
- Doyle MP, Hoekstra JW. Oxidation of nitrogen oxides by bound dioxygen in hemoproteins. J Inorg Biochem. 1981;14:351–358. S0162-0134(00)80291-3 [pii]
- Eich RF, et al. Mechanism of NO-induced oxidation of myoglobin and hemoglobin. Biochemistry. 1996;35:6976–6983. doi: 10.1021/bi960442gbi960442g[pii].
- Sharma VS, Traylor TG, Gardiner R, Mizukami H. Reaction of nitric oxide with heme proteins and model compounds of hemoglobin. Biochemistry. 1987;26:3837–3843.
- Tejero J, et al. Low NO concentration-dependence of the reductive nitrosylation reaction of hemoglobin. J Biol Chem. 2012 doi: 10.1074/jbc.M111.298927. M111.298927 [pii]
- Angelo M, Singel DJ, Stamler JS. An S-nitrosothiol (SNO) synthase function of hemoglobin that utilizes nitrite as a substrate. Proc Natl Acad Sci U S A. 2006;103:8366–8371. doi: 10.1073/pnas.0600942103. 0600942103 [pii]
- Lee E, Kariya K. Propylthiouracil, a selective inhibitor of NADH-cytochrome b5 reductase. FEBS Lett. 1986;209:49–51. 0014-5793(86)81082-1 [pii]
- Lam YH, Tang MH. Middle cerebral artery Doppler study in fetuses with homozygous alpha-thalassaemia-1 at 12-13 weeks of gestation. Prenat Diagn. 2002;22:56–58. doi: 10.1002/pd.237[pii].
- Fregly MJ, Hood CI. Physiologic and anatomic effects of prophylthiouracil on normal and hypertensive rats. Circ Res. 1959;7:486–496.
- Heberlein KR, et al. Plasminogen activator inhibitor-1 regulates myoendothelial junction formation. Circ Res. 2010;106:1092–1102. doi: 10.1161/CIRCRESAHA.109.215723. CIRCRESAHA.109.215723 [pii]
- Davalos A, et al. Quantitative proteomics of caveolin-1-regulated proteins: characterization of polymerase i and transcript release factor/CAVIN-1 IN endothelial cells. Mol Cell Proteomics. 2010;9:2109–2124. doi: 10.1074/mcp.M110.001289. M110.001289 [pii]
Source: PubMed