title: Enzymatic function of hemoglobin as a nitrite reductase that produces NO under allosteric control.
abstract: Hypoxic vasodilation is a fundamental, highly conserved physiological response that requires oxygen and/or pH sensing coupled to vasodilation. While this process was first characterized more than 80 years ago, the precise identity and mechanism of the oxygen sensor and mediators of vasodilation remain uncertain. In support of a possible role for hemoglobin (Hb) as a sensor and effector of hypoxic vasodilation, here we show biochemical evidence that Hb exhibits enzymatic behavior as a nitrite reductase, with maximal NO generation rates occurring near the oxy-to-deoxy (R-to-T) allosteric structural transition of the protein. The observed rate of nitrite reduction by Hb deviates from second-order kinetics, and sigmoidal reaction progress is determined by a balance between 2 opposing chemistries of the heme in the R (oxygenated conformation) and T (deoxygenated conformation) allosteric quaternary structures of the Hb tetramer--the greater reductive potential of deoxyheme in the R state tetramer and the number of unligated deoxyheme sites necessary for nitrite binding, which are more plentiful in the T state tetramer. These opposing chemistries result in a maximal nitrite reduction rate when Hb is 40-60% saturated with oxygen (near the Hb P50), an apparent ideal set point for hypoxia-responsive NO generation. These data suggest that the oxygen sensor for hypoxic vasodilation is determined by Hb oxygen saturation and quaternary structure and that the nitrite reductase activity of Hb generates NO gas under allosteric and pH control.
authors.affiliation: Vascular Therapeutics Section, Cardiovascular Branch, National Heart, Lung and Blood Institute, and Laboratory of Chemical Biology, National Institute of Diabetes, Digestive and Kidney Diseases, NIH, Bethesda, Maryland 20892-1662, USA.
authors.forename: Zhi
authors.initials: Z
authors.lastname: Huang
authors.forename: Sruti
authors.initials: S
authors.lastname: Shiva
authors.forename: Daniel B
authors.initials: DB
authors.lastname: Kim-Shapiro
authors.forename: Rakesh P
authors.initials: RP
authors.lastname: Patel
authors.forename: Lorna A
authors.initials: LA
authors.lastname: Ringwood
authors.forename: Cynthia E
authors.initials: CE
authors.lastname: Irby
authors.forename: Kris T
authors.initials: KT
authors.lastname: Huang
authors.forename: Chien
authors.initials: C
authors.lastname: Ho
authors.forename: Neil
authors.initials: N
authors.lastname: Hogg
authors.forename: Alan N
authors.initials: AN
authors.lastname: Schechter
authors.forename: Mark T
authors.initials: MT
authors.lastname: Gladwin
mesh_terms: D000494:Allosteric Regulation / Q000235:genetics; D000495:Allosteric Site / Q000235:genetics; D000818:Animals; D006454:Hemoglobins / Q000737:chemistry* / Q000235:genetics; D006736:Horses; D006801:Humans; D006863:Hydrogen-Ion Concentration; D000860:Hypoxia / Q000201:enzymology; D007700:Kinetics; D009569:Nitric Oxide / Q000737:chemistry*; D009572:Nitrite Reductases / Q000737:chemistry*; D009573:Nitrites / Q000737:chemistry*; D010084:Oxidation-Reduction; D017434:Protein Structure, Tertiary; D014664:Vasodilation
chemical_list: D006454:Hemoglobins; D009573:Nitrites; D009569:Nitric Oxide; D009572:Nitrite Reductases