The carboxyl terminus of the Saccharomyces cerevisiae succinate dehydrogenase membrane subunit, SDH4p, is necessary for ubiquinone reduction and enzyme stability.
The succinate dehydrogenase (SDH) of Saccharomyces cerevisiae is composed of four nonidentical subunits encoded by the nuclear genes SDH1, SDH2, SDH3, and SDH4. The hydrophilic subunits, SDH1p and SDH2p, comprise the catalytic domain involved in succinate oxidation. They are anchored to the inner mitochondrial membrane by two small, hydrophobic subunits, SDH3p and SDH4p, which are required for electron transfer and ubiquinone reduction. Comparison of the deduced primary sequence of the yeast SDH4p subunit to SDH4p subunits from other species reveals the presence of an unusual 25-30 amino acid carboxyl-terminal extension following the last predicted transmembrane domain. The extension is predicted to be on the cytoplasmic side of the inner mitochondrial membrane. To investigate the extension's function, three truncations were created and characterized. The results reveal that the carboxyl-terminal extension is necessary for respiration and growth on nonfermentable carbon sources, for ubiquinone reduction, and for enzyme stability. Combined with inhibitor studies using a ubiquinone analog, our results suggest that the extension and more specifically, residues 128-135 are involved in the formation of a ubiquinone binding site. Our findings support a two-ubiquinone binding site model for the S. cerevisiae SDH.
Oyedotun
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KS
Medical Research Council of Canada Group in the Molecular Biology of Membranes, Department of Biochemistry, University of Alberta, Edmonton, Alberta T6G 2H7, Canada.
Lemire
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BD
D019297:2,4-Dinitrophenol / Q000031:analogs & derivatives / Q000494:pharmacology; D000595:Amino Acid Sequence; D004795:Enzyme Stability; D005656:Fungal Proteins / Q000737:chemistry*; D007527:Isoenzymes / Q000737:chemistry*; D007700:Kinetics; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed; D010084:Oxidation-Reduction; D017433:Protein Structure, Secondary; D011809:Quinones / Q000378:metabolism; D012441:Saccharomyces cerevisiae / Q000201:enzymology*; D016415:Sequence Alignment; D017384:Sequence Deletion; D013385:Succinate Dehydrogenase / Q000737:chemistry*; D014451:Ubiquinone / Q000378:metabolism*
D005656:Fungal Proteins; D007527:Isoenzymes; D011809:Quinones; D014451:Ubiquinone; D013385:Succinate Dehydrogenase; D019297:2,4-Dinitrophenol; C018527:dinoseb