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A matrix-located processing peptidase of plant mitochondria

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Abstract

Nuclear-encoded mitochondrial precursor proteins are proteolytically processed inside the mitochondrion after import. The general mitochondrial processing activity in plant mitochondria has been shown to be integrated into the cytochrome bc1 complex of the respiratory chain. Here we investigate the occurrence of an additional, matrix-located processing activity by incubation of the precursors of the soybean mitochondrial proteins, alternative oxidase, the FAd subunit of the ATP synthetase and the tobacco F1β subunit of the ATP synthase, with the membrane and soluble components of mitochondria isolated from soybean cotyledons and spinach leaves. A matrix-located peptidase specifically processed the precursors to the predicted mature form in a reaction which was sensitive to orthophenanthroline, a characteristic inhibitor of mitochondrial processing peptidase (MPP). The specificity of the matrix peptidase was illustrated by the inhibition of processing of the alternative oxidase precursor in both soybean and spinach matrix extracts upon altering a single amino acid residue in the targeting presequence (-2 Arg to Gly). Additionally, there was no evidence for general proteolysis of precursor proteins incubated with the matrix. The purity of the matrix fractions was ascertained by spectrophotometric and immunological analyses. The results demonstrate that there is a specific processing activity in the matrix of soybean and spinach in addition to the previously well characterized membrane-bound MPP integrated into the cytochrome bc1 complex of the respiratory chain.

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Authors and Affiliations

  1. Department of Biochemistry, Arrhenius laboratories for Natural Sciences, Stockholm University, Stockholm, 106 91, Sweden

    Cristina Szigyarto, Carina Knorpp & Elzbieta Glaser

  2. Biochemistry Department, University of Western Australia, Nedlands, 6907, Perth, Western Australia, Australia

    Patrick Dessi & James Whelan

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

    M. Kathleen Smith & David A. Day

  4. Department of Botany, University College Dublin, Belfield, Donnybrook, Dublin, 4, Ireland

    Matthew A. Harmey

Authors
  1. Cristina Szigyarto
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  2. Patrick Dessi
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  3. M. Kathleen Smith
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  4. Carina Knorpp
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  5. Matthew A. Harmey
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  6. David A. Day
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  7. Elzbieta Glaser
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  8. James Whelan
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Szigyarto, C., Dessi, P., Smith, M.K. et al. A matrix-located processing peptidase of plant mitochondria. Plant Mol Biol 36, 171–181 (1998). https://doi.org/10.1023/A:1005977716814

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