title: Lucigenin and coelenterazine as superoxide probes in mitochondrial and bacterial membranes.
abstract: 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.
authors.affiliation: Department of Medical Biochemistry and Molecular Biology, University of Oulu, Box 5000, FIN-90014 Oulu, Finland.
authors.forename: Marko
authors.initials: M
authors.lastname: Kervinen
authors.forename: Jukka
authors.initials: J
authors.lastname: Pätsi
authors.forename: Moshe
authors.initials: M
authors.lastname: Finel
authors.forename: Ilmo E
authors.initials: IE
authors.lastname: Hassinen
mesh_terms: 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
chemical_list: 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