title: Stigmatellin induces reduction of iron-sulfur protein in the oxidized cytochrome bc1 complex.
abstract: Stigmatellin, a Q(P) site inhibitor, inhibits electron transfer from iron-sulfur protein (ISP) to cytochrome c1 in the bc1 complex. Stigmatellin raises the midpoint potential of ISP from 290 mV to 540 mV. The binding of stigmatellin to the fully oxidized complex, oxidized completely by catalytic amounts of cytochrome c oxidase and cytochrome c, results in ISP reduction. The extent of ISP reduction is proportional to the amount of inhibitor used and reaches a maximum when the ratio of inhibitor to enzyme complex reaches unity. A g = 2.005 EPR peak, characteristic of an organic free radical, is also observed when stigmatellin is added to the oxidized complex, and its signal intensity depends on the amount of stigmatellin. Addition of ferricyanide, a strong oxidant, to the oxidized complex also generates a g = 2.005 EPR peak that is oxidant concentration-dependent. Oxygen radicals are generated when stigmatellin is added to the oxidized complex in the absence of the exogenous substrate, ubiquinol. The amount of oxygen radical formed is proportional to the amount of stigmatellin added. Oxygen radicals are not generated when stigmatellin is added to a mutant bc1 complex lacking the Rieske iron-sulfur cluster. Based on these results, it is proposed that ISP becomes a strong oxidant upon stigmatellin binding, extracting electrons from an organic compound, likely an amino acid residue. This results in the reduction of ISP and generation of organic radicals.
authors.affiliation: Department of Biochemistry and Molecular Biology, Oklahoma State University, Stillwater, Oklahoma 74078, USA.
authors.forename: Buddha
authors.initials: B
authors.lastname: Gurung
authors.forename: Linda
authors.initials: L
authors.lastname: Yu
authors.forename: Chang-An
authors.initials: CA
authors.lastname: Yu
mesh_terms: D000818:Animals; D000900:Anti-Bacterial Agents / Q000494:pharmacology*; D000458:Cyanobacteria / Q000378:metabolism; D004578:Electron Spin Resonance Spectroscopy; D014450:Electron Transport Complex III / Q000737:chemistry*; D004583:Electrons; D005292:Ferricyanides / Q000494:pharmacology; D005786:Gene Expression Regulation; D006736:Horses; D007506:Iron-Sulfur Proteins / Q000378:metabolism*; D008954:Models, Biological; D016877:Oxidants / Q000737:chemistry; D010100:Oxygen / Q000737:chemistry*; D011090:Polyenes / Q000494:pharmacology; D013481:Superoxides / Q000737:chemistry
chemical_list: D000900:Anti-Bacterial Agents; D005292:Ferricyanides; D007506:Iron-Sulfur Proteins; D016877:Oxidants; D011090:Polyenes; D013481:Superoxides; C007931:hexacyanoferrate III; C041573:stigmatellin; D014450:Electron Transport Complex III; D010100:Oxygen