title: The reactivity of alpha-hydroxyhaem and verdohaem bound to haem oxygenase-1 to dioxygen and sodium dithionite.
abstract: Recently we have shown that ferric alpha-hydroxyhaem bound to haem oxygenase-1 can be converted to ferrous verdohaem by approximately an equimolar amount of O2 in the absence of exogenous electrons [Sakamoto, H., Omata, Y., Palmer, G., and Noguchi, M. (1999) J. Biol. Chem.274, 18196-18200]. Contrary to those results, other studies have claimed that the conversion requires both O2 and an electron. More recently, Migita et al. have reported that the major reaction product of ferric alpha-hydroxyhaem with O2 is a ferric porphyrin cation radical that can be converted to ferrous alpha-hydroxyhaem with sodium dithionite [Migita, C. T., Fujii, H., Matera, K. M., Takahashi, S., Zhou, H., and Yoshida, T. (1999) Biochim. Biophys. Acta1432, 203-213]. To clarify the reason(s) for the discrepancy, we compared the reactions; i.e. alpha-hydroxyhaem to verdohaem and verdohaem to biliverdin, under various conditions as well as according to the procedures of Migita. We find that complex formation of alpha-hydroxyhaem with haem oxygenase may be small and a substantial amount of free alpha-hydroxyhaem may remain, depending on the reconstitution conditions; this could lead to a misinterpretation of the experimental results. We also find that ferrous verdohaem appears to be air-sensitive and is therefore easily converted to a further oxidized species with excess O2. Finally, we find that dithionite seems to be inappropriate for investigating the haem oxygenase reaction, because it reduces ferrous verdohaem to a further reduced species that has not been seen in the haem degradation system driven by NADPH-cytochrome P450 reductase.
authors.affiliation: Department of Medical Biochemistry, Kurume University School of Medicine, Japan.
authors.forename: Hiroshi
authors.initials: H
authors.lastname: Sakamoto
authors.forename: Yoshiaki
authors.initials: Y
authors.lastname: Omata
authors.forename: Shunsuke
authors.initials: S
authors.lastname: Hayashi
authors.forename: Saori
authors.initials: S
authors.lastname: Harada
authors.forename: Graham
authors.initials: G
authors.lastname: Palmer
authors.forename: Masato
authors.initials: M
authors.lastname: Noguchi
mesh_terms: D002248:Carbon Monoxide / Q000737:chemistry; D004227:Dithionite / Q000737:chemistry*; D004578:Electron Spin Resonance Spectroscopy; D006418:Heme / Q000031:analogs & derivatives* / Q000737:chemistry*; D006419:Heme Oxygenase (Decyclizing) / Q000737:chemistry*; D051547:Heme Oxygenase-1; D046911:Macromolecular Substances; D009251:NADPH-Ferrihemoprotein Reductase / Q000737:chemistry; D010084:Oxidation-Reduction; D010100:Oxygen / Q000737:chemistry*; D011166:Porphyrins / Q000737:chemistry; D013053:Spectrophotometry; D013055:Spectrophotometry, Infrared
chemical_list: D046911:Macromolecular Substances; D011166:Porphyrins; C099546:alpha-hydroxyheme; C099547:verdoheme; D004227:Dithionite; D006418:Heme; D002248:Carbon Monoxide; D006419:Heme Oxygenase (Decyclizing); D051547:Heme Oxygenase-1; D009251:NADPH-Ferrihemoprotein Reductase; D010100:Oxygen