Gas chromatographic differentiation between myeloperoxidase activity and Fenton-type oxidants.
Several pathological situations are characterized by the production of reactive oxygen species (ROS), whereby different sources such as activated leukocytes and xanthine oxidase seem to be mainly responsible. The contribution of immigrating activated neutrophils to symptom development during inflammatory processes or after reperfusion of ischemic tissues is a matter of continuing discussion. We present a simple method for the differentiation between oxygen activating reactions in which neutrophil-derived myeloperoxidase is involved. The method is based on the gas chromatographic detection of ethylene, which is formed by the reaction of alpha-keto-gamma-methylthiobutyric acid (KMB) or 1-amino-cyclopropane-1-carboxylic acid (ACC) with ROS. In the presence of OH-radical-type oxidants, only KMB yields ethylene whereas ACC is fragmented by myeloperoxidase-derived species (OCl-, chloramines). The amounts of ethylene may be used as an indicator for the relative contribution of Fenton-type or myeloperoxidase-catalyzed reactions.
von Kruedener
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Lehrstuhl für Phytopathologie, Labor für Angewandte Biochemie, Technische Universität München, Weihenstephan, Germany.
Schempp
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H
Elstner
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EF
D000596:Amino Acids / Q000737:chemistry; D000598:Amino Acids, Cyclic; D002849:Chromatography, Gas; D005030:Ethylenes / Q000737:chemistry; D005609:Free Radicals; D006861:Hydrogen Peroxide; D006863:Hydrogen-Ion Concentration; D007501:Iron; D007700:Kinetics; D008715:Methionine / Q000031:analogs & derivatives / Q000737:chemistry; D016877:Oxidants; D010084:Oxidation-Reduction; D009195:Peroxidase / Q000378:metabolism*; D017382:Reactive Oxygen Species / Q000737:chemistry; D019820:Xanthine; D014969:Xanthine Oxidase / Q000378:metabolism; D014970:Xanthines / Q000378:metabolism
D000596:Amino Acids; D000598:Amino Acids, Cyclic; D005030:Ethylenes; C045076:Fenton's reagent; D005609:Free Radicals; D016877:Oxidants; D017382:Reactive Oxygen Species; D014970:Xanthines; D019820:Xanthine; C023863:1-aminocyclopropane-1-carboxylic acid; C014632:2-keto-4-methylthiobutyric acid; C036216:ethylene; D008715:Methionine; D006861:Hydrogen Peroxide; D007501:Iron; D009195:Peroxidase; D014969:Xanthine Oxidase