Structure/function relationships in mitochondrial cytochrome b revealed by the kinetic and circular dichroic properties of two yeast inhibitor-resistant mutants.
The kinetic and circular dichroic properties of two yeast mutants that are resistant towards specific inhibitors of the mitochondrial cytochrome bc1 complex have been characterized. Both of these mutants have an altered cytochrome b gene in which aromatic residues are exchanged with non-polar residues in a highly conserved region of the protein. The mutant resistant to myxothiazol and mucidin that contains the substitution Phe129----Leu is not greatly affected either in its ubiquinol:cytochrome c reductase or in the spectral properties of cytochrome b. On the other hand, the mutant resistant to stigmatellin that contains the substitution Ile147----Phe shows a large decrease of the catalytic efficiency for ubiquinol and of the maximal turnover of its reductase activity. This stigmatellin mutant also shows an altered circular-dichroic spectrum of the low-potential haem of cytochrome b. This study provides biochemical and biophysical information for identifying a region in mitochondrial cytochrome b that may fulfill a crucial role in the binding of ubiquinol to the bc1 complex. The results are discussed also in terms of the structural model of cytochrome b having a core of four transmembrane helices.
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Laboratore de Genetique Microbienne, Université Catholique de Louvain-la-Neuve, Belgium.
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Colson
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Degli Esposti
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D000595:Amino Acid Sequence; D002942:Circular Dichroism; D003573:Cytochrome b Group / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D014450:Electron Transport Complex III / Q000037:antagonists & inhibitors / Q000378:metabolism; D005231:Fatty Acids, Unsaturated / Q000494:pharmacology; D007700:Kinetics; D008689:Methacrylates; D008969:Molecular Sequence Data; D009154:Mutation; D011090:Polyenes / Q000494:pharmacology; D011487:Protein Conformation; D012441:Saccharomyces cerevisiae / Q000378:metabolism*; D000073739:Strobilurins; D013844:Thiazoles / Q000494:pharmacology
D003573:Cytochrome b Group; D005231:Fatty Acids, Unsaturated; D008689:Methacrylates; D011090:Polyenes; D000073739:Strobilurins; D013844:Thiazoles; C008556:mucidin; C030517:myxothiazol; C041573:stigmatellin; D014450:Electron Transport Complex III