Article ID Journal Published Year Pages File Type
10768218 Biochemical and Biophysical Research Communications 2005 6 Pages PDF
Abstract
O2-dependent reactions of the ferric and ferrous forms of α-hydroxyheme complexed with water-soluble rat heme oxygenase-1 were examined by rapid-scan stopped-flow measurements. Ferric α-hydroxyheme reacted with O2 to form ferric verdoheme with an O2-dependent rate constant of 4 × 105 M−1 s−1 at pH 7.4 and 9.0. A decrease of the rate constant to 2.8 × 105 M−1 s−1 at pH 6.5 indicates that the reaction proceeds by direct attack of O2 on the π-neutral radical form of α-hydroxyheme, which is generated by deprotonation of the α-hydroxy group. The reaction of ferrous α-hydroxyheme with O2 yielded ferrous verdoheme in a biphasic fashion involving a new intermediate having absorption maxima at 415 and 815 nm. The rate constants for this two-step reaction were 68 and 145 s−1. These results show that conversion of α-hydroxyheme to verdoheme is much faster than the reduction of coordinated iron (<1 s−1) under physiological conditions [Y. Liu, P.R. Ortiz de Montellano, Reaction intermediates and single turnover rate constants for the oxidation of heme by human heme oxygenase-1, J. Biol. Chem. 275 (2000) 5297-5307], suggesting that, in vivo, the conversion of ferric α-hydroxyheme to ferric verdoheme precedes the reduction of ferric α-hydroxyheme.
Related Topics
Life Sciences Biochemistry, Genetics and Molecular Biology Biochemistry
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