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The general mitochondrial processing peptidase from wheat is integrated into the cytochrome bc 1-complex of the respiratory chain

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Abstract

The bc 1-complex (EC 1.10.2.2.) from Triticum aestivum L. was purified by cytochrome-c affinity chromatography and gel filtration using either etiolated seedlings or wheat-germ extract as starting material. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis of the isolated enzyme revealed ten bands, which were analysed by immunoblotting and direct amino-acid sequencing. The enzyme from wheat is the first bc 1-complex that is reported to contain four core proteins (55.5, 55.0, 51.5 and 51.0 kDa). In addition, the wheat bc 1-complex comprises cytochrome b (35 kDa), cytochrome c 1 (33 kDa) the “Rieske” iron-sulphur protein (25 kDa) and three small subunits < 15 kDa. This composition differs from the one reported in fungi, mammals and potato. Partial sequence determination of the large subunits suggests that the 55.5 and 55.0-kDa-proteins represent the β-subunit of the general mitochondrial processing peptidase, and the 51.5 and 51.0-kDa proteins the α-subunit of this enzyme. The bc 1-complex from wheat efficiently processes mitochondrial precursor proteins as shown in an in-vitro processing assay. In control experiments the isolated bc 1-complexes from potato, yeast, Neurospora and beef, all purified by the same isolation procedure, were also tested for processing activity. Only the protein complexes from plants contain the general mitochondrial processing peptidase. The composition of the wheat bc 1-complex sheds new light on the co-evolution of the processing peptidase and the middle segment of the respiratory chain.

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Abbreviations

MPP:

mitochondrial processing peptidase

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Authors and Affiliations

  1. Institut für Genbiologische Forschung Berlin GmbH, Ihnestr. 63, D-14195, Berlin, Germany

    Hans-Peter Braun, Michael Emmermann & Udo K. Schmitz

  2. Applied Biosystems GmbH, Brunnenweg 13, D-64331, Weiterstadt, Germany

    Volker Kruft

  3. Institut für Biochemie, Universität Düsseldorf, Universitätsstr. 1, D-40225, Düsseldorf, Germany

    Martin Bödicker

Authors
  1. Hans-Peter Braun
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  2. Michael Emmermann
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  3. Volker Kruft
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  4. Martin Bödicker
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  5. Udo K. Schmitz
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Corresponding author

Correspondence to Udo K. Schmitz.

Additional information

We wish to thank Prof. G. Schatz, Biozentrum Basel, Switzerland and Prof. H. Weiss, Universität Düsseldorf, Germany for providing antibodies against the repiratory subunits of the bc 1-complex from yeast and Neurospora and to H. Mentzel, A. Leisse, R. Breitfeld and B. Hidde for excellent technical assistance. Thanks are also due to Prof. M. Boutry, Université de Louvaine-la-Neuve, Belgium for providing a plasmid containing the β-subunit of ATPase from tobacco. This research was supported by the Deutsche Forschungsgemeinschalft and the Bundesministerium für Forschung und Technologie.

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Braun, HP., Emmermann, M., Kruft, V. et al. The general mitochondrial processing peptidase from wheat is integrated into the cytochrome bc 1-complex of the respiratory chain. Planta 195, 396–402 (1995). https://doi.org/10.1007/BF00202597

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