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Studies on the import and processing of the alternative oxidase precursor by isolated soybean mitochondria

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Abstract

Import of the synthetic precursor of the alternative oxidase from soybean was shown to be dependent on a membrane potential and ATP. The membrane potential in soybean mitochondria may be formed either by respiration through the cytochrome pathway, or through the alternative oxidase pathway with NAD+-linked substrates. Import of the alternative oxidase precursor in the presence of succinate as respiratory substrate was inhibited by KCN. Import in the presence of malate was insensitive to KCN and SHAM added separately, but was inhibited by KCN and SHAM added together (inhibitors of the cytochrome and alternative oxidases respectively). Import of the alternative oxidase was accompanied by processing of the precursor to a single 32 kDa product in both cotyledon and root mitochondria. This product had a different mobility than the two alternative oxidase bands detected by immunological means (34 and 36 kDa), suggesting that the enzyme had been modified in situ. When the cDNA clone of the alternative oxidase was modified by a single mutation (−2 Arg changed to −2 Gly), the processing of the precursor was inhibited.

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Author notes

  1. James Whelan

    Present address: Department of Biochemistry, University of Western Australia, 6009, Nedlands, Western Australia, Australia

Authors and Affiliations

  1. Division of Biochemistry and Molecular Biology, School of Life Sciences, Australian National University, 0200, Canberra, ACT, Australia

    James Whelan & David A. Day

  2. Department of Biochemistry, Arrhenius Laboratories for Natural Science, Stockholm University, S-106 91, Stockholm, Sweden

    Marie Hugosson & Elzbieta Glaser

Authors
  1. James Whelan
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  2. Marie Hugosson
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  3. Elzbieta Glaser
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  4. David A. Day
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Whelan, J., Hugosson, M., Glaser, E. et al. Studies on the import and processing of the alternative oxidase precursor by isolated soybean mitochondria. Plant Mol Biol 27, 769–778 (1995). https://doi.org/10.1007/BF00020229

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