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

Mol Gen Genet. 1997 Apr 28;254(4):365-71. doi: 10.1007/s004380050428.

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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Cell Fractionation
  • Centrifugation, Density Gradient
  • Electrophoresis, Gel, Two-Dimensional
  • Fabaceae / chemistry
  • Fabaceae / genetics*
  • Genome, Plant*
  • Mitochondria / genetics*
  • Mitochondria / ultrastructure
  • Molecular Sequence Data
  • Plant Proteins / chemistry
  • Plant Proteins / genetics*
  • Plants, Medicinal*
  • Ribosomal Proteins / analysis
  • Ribosomal Proteins / chemistry
  • Ribosomal Proteins / genetics*
  • Ribosomes / chemistry*
  • Sequence Alignment

Substances

  • Plant Proteins
  • Ribosomal Proteins
  • ribosomal protein S10
  • ribosomal protein S12