Role of membrane-bound and free polyribosomes in the synthesis of cytochrome c in rat liver.
The functional distinction of membrane-bound and free polyribosomes for the synthesis of exportable and non-exportable proteins respectively is not so strict as was initially thought, and it was therefore decided to investigate their relative contribution to the elaboration of an internal protein integrated into a cell structure. Cytochrome c was chosen as an example of a soluble mitochondrial protein, and the incorporation of [(14)C]leucine and delta-amino[(14)C]laevulinate into the molecule was studied by using different ribosomal preparations from regenerating rat liver. A new procedure was devised for the purification of cytochrome c, based on ion-exchange chromatography combined with sodium dodecyl sulphate-polyacrylamide-gel electrophoresis. In spite of cytochrome c being a non-exportable protein, the membrane-bound polyribosomes were at least as active as the free ribosomes in the synthesis in vitro of the apoprotein and the haem moiety. The detergent-treated ribosomes could also effect the synthesis of cytochrome c, although at a lower rate. Since in liver more than two-thirds of the ribosomes are bound to the endoplasmic-reticulum membranes, it is considered that in vivo they are responsible for the synthesis of most of the cytochrome c content of the cell. This suggests that in secretory tissues the endoplasmic reticulum plays a predominant role in mitochondrial biogenesis, although free ribosomes may participate in the partial turnover of some parts of the organelle. The hypothesis on the functional specialization of the different kinds of ribosomes was therefore modified to account for their parallel intervention in the synthesis of proteins associated with membranous structures.
D000818:Animals; D002250:Carbon Radioisotopes; D002498:Centrifugation; D002852:Chromatography, Ion Exchange; D003574:Cytochrome c Group / Q000096:biosynthesis*; D003956:Dialysis; D004591:Electrophoresis, Polyacrylamide Gel; D004721:Endoplasmic Reticulum; D066298:In Vitro Techniques; D007930:Leucine / Q000378:metabolism; D007982:Levulinic Acids / Q000378:metabolism; D008099:Liver / Q000378:metabolism*; D008566:Membranes; D008930:Mitochondria, Liver / Q000378:metabolism; D011132:Polyribosomes / Q000378:metabolism*; D011485:Protein Binding; D051381:Rats; D012270:Ribosomes; D014465:Ultrasonics
D002250:Carbon Radioisotopes; D003574:Cytochrome c Group; D007982:Levulinic Acids; D007930:Leucine