title: Diphenyleneiodonium inhibits reduction of iron-sulfur clusters in the mitochondrial NADH-ubiquinone oxidoreductase (Complex I).
abstract: Diphenyleneiodonium (DPI) inhibits the mitochondrial NADH-ubiquinone oxidoreductase (Complex I) on the substrate side of the Fe-S clusters. In the inhibited NADH-supplemented state all of the Fe-S clusters are oxidized, whereas the reduced minus oxidized difference spectrum of the protein-bound FMN can be visualized. It is characterized by troughs at 370 and 450 nm and a small increase of absorbance in the 500-700-nm region. DPI probably reacts irreversibly with FMN, because oxidation of FMN is blocked even after its extraction from the enzyme. Inhibition requires preincubation of enzyme in the presence of NADH and DPI. The lower the NADH/NAD+ ratio or the pH, or the higher the NAD+/DPI ratio, the more DPI is required for inhibition. NAD+ and DPI apparently compete for a common site. Both ubiquinone and dichlorophenolindophenol reductase activities are fully blocked by DPI, whereas the ferricyanide reductase activity is inhibited by 75%. Similar results were found with Complex I and two rotenone-insensitive preparations, subcomplex I lambda and the flavoprotein fraction. DPI also inhibits NADH oxidation by bacterial NADH-ubiquinone oxidoreductase-1 (NDH-1) in membranes of Paracoccus denitrificans and Escherichia coli.
authors.affiliation: Department of Medical Chemistry, University of Helsinki, Finland.
authors.forename: A
authors.initials: A
authors.lastname: Majander
authors.forename: M
authors.initials: M
authors.lastname: Finel
authors.forename: M
authors.initials: M
authors.lastname: Wikström
mesh_terms: D004578:Electron Spin Resonance Spectroscopy; D004926:Escherichia coli / Q000201:enzymology; D005486:Flavin Mononucleotide / Q000737:chemistry; D007506:Iron-Sulfur Proteins / Q000187:drug effects / Q000378:metabolism*; D008928:Mitochondria / Q000201:enzymology*; D016660:NAD(P)H Dehydrogenase (Quinone) / Q000037:antagonists & inhibitors / Q000737:chemistry / Q000378:metabolism*; D009861:Onium Compounds / Q000494:pharmacology*; D010084:Oxidation-Reduction; D010231:Paracoccus denitrificans / Q000201:enzymology
chemical_list: D007506:Iron-Sulfur Proteins; D009861:Onium Compounds; C007517:diphenyleneiodonium; D005486:Flavin Mononucleotide; D016660:NAD(P)H Dehydrogenase (Quinone)