Preparation and some properties of rat skeletal-muscle polyribosomes

Abstract

1. A method is described for the sucrose-gradient sedimentation analysis of ribosomes in a post-mitochondrial supernatant of rat skeletal muscle. 2. An essential feature of the method involves the use of buffer of ionic strength 0·3 for homogenization of the muscle tissue. 3. Polyribosomes can be prepared by precipitation from post-mitochondrial supernatant of skeletal muscle by adjustment of the potassium chloride content of the medium. These polyribosomes stimulate cell-free amino acid incorporation in vitro in an energy-dependent system. 4. Ribosome aggregates of uniform size distribution can be obtained by adjustment of the ionic strength of the post-mitochondrial supernatant, followed by differential sucrose-gradient centrifugation. 5. In vivo, rat skeletal-muscle polyribosomes became labelled by ¹⁴C-labelled amino acid within 15min., and radioactivity was associated with the light ribosome species within 45min. 6. Electron microscopy of the polyribosomes revealed aggregations containing more than 40 single ribosomes.