Hemoglobin: A mechanism for the generation of hydroxyl radicals

• Published in: Free radical biology & medicine Vol. 20; no. 7; pp. 985 - 989
• Main Authors: Van Dyke, Bruce R., Saltman, Paul
• Format: Journal Article
• Language: English
• Published: United States Elsevier Inc 1996
• Subjects: Amino Acids - chemistry; Buffers; Carbonyl formation; Electron Transport; Free radical scavengers; Free radicals; Hemoglobin; Hemoglobins - chemistry; Hydroxybenzoates - metabolism; Hydroxyl radical; Hydroxyl Radical - metabolism; Oxidation-Reduction; Redox metals; Carbonyl formation; Hemoglobin; Free radicals; Redox metals; Hydroxyl radical; Free radical scavengers
• ISSN: 0891-5849; 1873-4596
• DOI: 10.1016/0891-5849(95)02186-8

Oxyhemoglobin (HbO 2) reduces Fe(III) NTA aerobically to become methemoglobin (metHb) and Fe(II)NTA. These conditions are favorable for the generation via Fenton chemistry of the hydroxyl radical that was measured by HPLC using salicylate as a probe. The levels of hydroxyl radicals generated are a function of both the percent metHb formed and the chemical nature of the buffer. The rates of formation of both metHb and hydroxyl radicals were dependent upon the concentration of Fe(III)NTA. Of the buffers tested, HEPES was the most effective scavenger of hydroxyl radicals while the other buffers scavenged in the order: HEPES > Tris > MOPS > NaCl ≈ unbuffered. The addition of catalase to remove H 20 2 or bathophenanthroline to chelate Fe(II) inhibited virtually all hydroxyl radical formation. Carbonyl formation from free radical oxidation of amino acids was found to be 0.1 mol/mol of hemoglobin. These experiments demonstrate the ability of hemoglobin to participate directly in the generation of hydroxyl radicals mediated by redox metals, and provide insight into potential oxidative damage from metals released into the blood during some pathologic disorders including iron overload.