Reactions of superoxide with myeloperoxidase.
When neutrophils ingest bacteria, they discharge superoxide and myeloperoxidase into phagosomes. Both are essential for killing of the phagocytosed micro-organisms. It is generally accepted that superoxide is a precursor of hydrogen peroxide which myeloperoxidase uses to oxidize chloride to hypochlorous acid. Previously, we demonstrated that superoxide modulates the chlorination activity of myeloperoxidase by reacting with its ferric and compound II redox states. In this investigation we used pulse radiolysis to determine kinetic parameters of superoxide reacting with redox forms of myeloperoxidase and used these data in a steady-state kinetic analysis. We provide evidence that superoxide reacts with compound I and compound III. Our estimates of the rate constants for the reaction of superoxide with compound I, compound II, and compound III are 5 x 10(6) M-1 s-1, 5.5 +/- 0.4 x 10(6) M-1 s-1, and 1.3 +/- 0.2 x 10(5) M-1 s-1, respectively. These reactions define new activities for myeloperoxidase. It will act as a superoxide dismutase when superoxide reacts consecutively with ferric myeloperoxidase and compound III. It will also act as a superoxidase by using hydrogen peroxide to oxidize superoxide via compound I and compound II. The favorable kinetics of these reactions indicate that, within the confines of a phagosome, superoxide will react with myeloperoxidase and affect the reactions it will catalyze. These interactions of superoxide and myeloperoxidase will have a major influence on the way neutrophils use oxygen to kill bacteria. Consequently, superoxide should be viewed as a cosubstrate that myeloperoxidase uses to elicit bacterial killing.
Kettle
Anthony J
AJ
Free Radical Research, Department of Pathology, University of Otago Christchurch, P.O. Box 4345 Christchurch, New Zealand. tony.kettle@chmeds.ac.nz
Anderson
Robert F
RF
Hampton
Mark B
MB
Winterbourn
Christine C
CC
D005290:Ferric Compounds / Q000378:metabolism; D006801:Humans; D007700:Kinetics; D009195:Peroxidase / Q000378:metabolism*; D011675:Pulse Radiolysis; D013482:Superoxide Dismutase / Q000378:metabolism; D013481:Superoxides / Q000737:chemistry*
D005290:Ferric Compounds; D013481:Superoxides; D009195:Peroxidase; D013482:Superoxide Dismutase