Partial purification of mitochondrial ribosomes from broad bean and identification of proteins encoded by the mitochondrial genome |
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Authors: | L Maffey H Degand and M Boutry |
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Institution: | (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.be, BE |
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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.
Received: 4 September 1996 / Accepted: 30 November 1996 |
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Keywords: | Plant mitochondria Ribosomes Mitochondrial translation products Broad bean Protein sequencing |
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