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Molecular and General Genetics MGG

, Volume 254, Issue 4, pp 365–371 | Cite as

Partial purification of mitochondrial ribosomes from broad bean and identification of proteins encoded by the mitochondrial genome

  • L. Maffey
  • H. Degand
  • M. Boutry
ORIGINAL PAPER

Abstract

An approach towards the identification at the protein level of the ribosomal proteins encoded by the mitochondrial genome of broad bean (Vicia faba) has been developed. After Triton X-100 treatment of isolated mitochondria, a fraction enriched in mitochondrial ribosomes was obtained by successive centrifugation, first onto a sucrose cushion, and then in a sucrose gradient. Mitochondrial translation products were labelled in isolated mitochondria with [35S]methionine and added to the enriched mitochondrial ribosomal proteins before separation by two-dimensional gel electrophoresis. Six spots, identified both by Coomassie blue staining and autoradiography, were analysed by protein micro-sequencing. Two of these were shown to correspond to ribosomal proteins S10 and S12. We conclude that these two proteins are encoded by the mitochondrial genome of broad bean and that the method described here can be used to identify other proteins encoded by the mitochondrial genome.

Key words Plant mitochondria   Ribosomes    Mitochondrial translation products   Broad bean    Protein sequencing 

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Copyright information

© Springer-Verlag Berlin Heidelberg 1997

Authors and Affiliations

  • L. Maffey
    • 1
  • H. Degand
    • 1
  • M. Boutry
    • 1
  1. 1.Unité de Biochimie Physiologique, Université catholique de Louvain, Place Croix du Sud 2-20, B-1348 Louvain-la-Neuve, Belgium tel. +32-10-473621 fax: +32-10-473872 e-mail boutry@fysa.ucl.ac.beBE

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