Kinetics of chlorination of monochlorodimedone by myeloperoxidase.
The phagocyte-derived enzyme myeloperoxidase has been recently implicated in the pathogenesis of atherosclerosis, because it catalyzes the reaction of hydrogen peroxide with chloride ions to give the highly toxic oxidant hypochlorous acid. The aim of this study was to determine the dependence of this reaction on the concentration of hydrogen peroxide and of the enzyme by means of the photometric monochlorodimedone assay. The initial rate of hypochlorous acid formation increased less than proportionally with increasing myeloperoxidase concentrations. Variation of the concentration of hydrogen peroxide had a biphasic effect, with an optimal concentration of hydrogen peroxide. Above this concentration enzyme destruction is apparently predominant. The progress curves of hypochlorous acid formation showed two distinct maxima. It was concluded that hypochlorous acid not only reacts with monochlorodimedone but also with the amino groups of myeloperoxidase to form intermediary chloramines that may further chlorinate monochlorodimedone. This was supported by the kinetics in the presence of the amino compound glycine, a competitive substrate for chlorination by hypochlorous acid. In the presence of high concentrations of glycine the progress curve rises continuously, yielding a greatly increased concentration of chlorinating species, either hypochlorous acid or chloramines. We concluded that glycine protects myeloperoxidase against hypochlorous acid-induced self-destruction.
Jerlich
A
A
Institute of Molecular Biology, Biochemistry, and Microbiology, University of Graz, Austria.
Tschabuschnig
S
S
Fabjan
J S
JS
Schaur
R J
RJ
D001667:Binding, Competitive; D003512:Cyclohexanones / Q000378:metabolism*; D005998:Glycine / Q000378:metabolism*; D006801:Humans; D006861:Hydrogen Peroxide / Q000378:metabolism; D006997:Hypochlorous Acid / Q000378:metabolism*; D007700:Kinetics; D009504:Neutrophils / Q000201:enzymology; D009195:Peroxidase / Q000378:metabolism*
D003512:Cyclohexanones; D006997:Hypochlorous Acid; C006991:chlorodimedone; D006861:Hydrogen Peroxide; D009195:Peroxidase; D005998:Glycine