Effects of neopterin-derivatives on H2O2-induced luminol chemiluminescence: mechanistic aspects.
Neopterin, 6-D-erythro-1',2',3'-trihydroxypropyl-pterin, and its dihydroform, 7,8-dihydro-neopterin, are synthesized by human monocytes/macrophages upon stimulation by interferon-gamma. In the presence of iron chelator complexes neopterin enhances hydrogen peroxide-induced luminol chemiluminescence at neutral or slightly alkaline pH (7.5). In contrast, 7,8-dihydroneopterin scavenges chemiluminescence independently from the pH value and iron. In this study, we explored in more detail the mechanism possibly involved: analysis of the reaction products shows that 7,8-dihydroneopterin is oxidized and degraded to 7,8-dihydroxanthopterin and xanthopterin, whereas the neopterin molecule is not chemically altered during the chemiluminescence reaction. Investigations of the neopterin-induced effect show that mannitol, a scavenger of hydroxyl radicals, does not alter the enhancing effect of neopterin. L-histidine, which scavenges singlet oxygen almost as effective as hydroxyl radicals, reduces the enhancing effect of neopterin. However, singlet oxygen was not detectable during the reaction by measuring monomol light emission (1270 nm). When replacing hydrogen peroxide by 3-morpholinosydnonimine, a generator of hydroxyl radicals, or naphthalene-endoperoxide, a generator of singlet oxygen, in the luminol chemiluminescence assay, neopterin shows no enhancing effect irrespective of the presence of iron-(III)-EDTA. The data suggest that neopterin enhances hydrogen peroxide-induced luminol chemiluminescence in the presence of iron-(III)-EDTA by formation of a catalytic complex that seems to favor the formation of oxygen intermediates which derive from hydrogen peroxide and react with luminol.
Murr
C
C
Institute for Medical Chemistry and Biochemistry, University of Innsbruck, Austria.
Baier-Bitterlich
G
G
Fuchs
D
D
Werner
E R
ER
Esterbauer
H
H
Pfleiderer
W
W
Wachter
H
H
D001708:Biopterins / Q000031:analogs & derivatives* / Q000096:biosynthesis / Q000494:pharmacology*; D002851:Chromatography, High Pressure Liquid; D004492:Edetic Acid; D005290:Ferric Compounds; D016166:Free Radical Scavengers; D006639:Histidine; D006801:Humans; D006861:Hydrogen Peroxide; D007371:Interferon-gamma / Q000494:pharmacology; D007502:Iron Chelating Agents; D007700:Kinetics; D008163:Luminescent Measurements; D008165:Luminol; D008264:Macrophages / Q000187:drug effects / Q000502:physiology; D009000:Monocytes / Q000187:drug effects / Q000502:physiology; D019798:Neopterin; D010100:Oxygen / Q000032:analysis; D011621:Pteridines / Q000032:analysis; D026082:Singlet Oxygen
D005290:Ferric Compounds; D016166:Free Radical Scavengers; D007502:Iron Chelating Agents; D011621:Pteridines; D026082:Singlet Oxygen; D001708:Biopterins; D006639:Histidine; D008165:Luminol; D019798:Neopterin; D007371:Interferon-gamma; D004492:Edetic Acid; D006861:Hydrogen Peroxide; C019179:Fe(III)-EDTA; D010100:Oxygen