Lucigenin and coelenterazine as superoxide probes in mitochondrial and bacterial membranes.
The chemiluminescent superoxide indicators lucigenin and coelenterazine were compared in rat liver submitochondrial particles and cytoplasmic membranes from Paracoccus denitrificans. Qualitative monitoring is possible with both probes, but quantitative work with lucigenin is hampered by its dependence on one-electron reduction before the photon-emitting reaction. Therefore, calibration of measurements on complex I, capable of efficient lucigenin prereduction with reduced nicotinamide adenine dinucleotide, against xanthine oxidase, which in the presence of hypoxanthine is not able to reduce the probe to a significant rate compared to complex I, may give results in error by one order of magnitude. Coelenterazine, although susceptible of storage-dependent high background chemiluminescence, does not require prereduction and is thus a more reliable probe.
Kervinen
Marko
M
Department of Medical Biochemistry and Molecular Biology, University of Oulu, Box 5000, FIN-90014 Oulu, Finland.
Pätsi
Jukka
J
Finel
Moshe
M
Hassinen
Ilmo E
IE
D000166:Acridines / Q000494:pharmacology*; D000818:Animals; D002462:Cell Membrane / Q000378:metabolism; D049454:Firefly Luciferin / Q000494:pharmacology*; D019271:Hypoxanthine / Q000378:metabolism; D007093:Imidazoles; D007425:Intracellular Membranes / Q000378:metabolism; D008163:Luminescent Measurements; D008930:Mitochondria, Liver / Q000187:drug effects / Q000201:enzymology*; D015335:Molecular Probes; D009247:NADH, NADPH Oxidoreductases / Q000378:metabolism; D010084:Oxidation-Reduction; D010231:Paracoccus denitrificans / Q000187:drug effects / Q000201:enzymology*; D011719:Pyrazines / Q000494:pharmacology*; D051381:Rats; D013481:Superoxides / Q000032:analysis / Q000378:metabolism*; D013997:Time Factors; D014969:Xanthine Oxidase / Q000494:pharmacology
D000166:Acridines; D007093:Imidazoles; D015335:Molecular Probes; D011719:Pyrazines; D013481:Superoxides; C033472:10,10'-dimethyl-9,9'-biacridinium; D019271:Hypoxanthine; C017144:coelenterazine; D049454:Firefly Luciferin; D014969:Xanthine Oxidase; D009247:NADH, NADPH Oxidoreductases