Organization of proteins in mammalian mitochondrial ribosomes. Accessibility to lactoperoxidase-catalyzed radioiodination.
To assess the relative exposure of individual ribosomal proteins (r-proteins) in the large and small subunits of the bovine mitochondrial ribosome, we used a double label iodination technique. Regions of r-proteins exposed in purified ribosomal subunits were labeled with 131I using the lactoperoxidase-catalyzed iodination system, and additional reactive groups available upon denaturing the r-proteins in urea were labeled with 125I using the chloramine-T mediated reaction. The ratio of 131I to 125I incorporated into individual proteins under these conditions is representative of the degree of exposure for each of the proteins in the subunits. In this manner, the r-proteins have been grouped into 3 classes depending on their degree of exposure: high exposure, intermediate exposure, and essentially buried. While both subunits have a few proteins in the "highly exposed" group, and a large number of proteins in the "intermediate exposure" group, only the large ribosomal subunit has an appreciable number of proteins which appear essentially buried. The more buried proteins may serve mainly structural roles, perhaps acting as "assembly proteins," since many from this group bind to ribosomal RNA. The more superficially disposed proteins may comprise binding sites for macromolecules that interact with ribosomes during protein synthesis, as well as stabilizing the association of the large and small subribosomal particles.
D000818:Animals; D002417:Cattle; D002700:Chloramines / Q000378:metabolism; D002712:Chlorides / Q000494:pharmacology; D004591:Electrophoresis, Polyacrylamide Gel; D007457:Iodine Radioisotopes / Q000378:metabolism*; D007784:Lactoperoxidase / Q000378:metabolism*; D008094:Lithium / Q000494:pharmacology; D018021:Lithium Chloride; D046911:Macromolecular Substances; D010544:Peroxidases / Q000378:metabolism*; D011489:Protein Denaturation; D012269:Ribosomal Proteins / Q000032:analysis*; D012270:Ribosomes / Q000032:analysis*; D013499:Surface Properties; D014105:Tosyl Compounds; D014508:Urea / Q000494:pharmacology
D002700:Chloramines; D002712:Chlorides; D007457:Iodine Radioisotopes; D046911:Macromolecular Substances; D012269:Ribosomal Proteins; D014105:Tosyl Compounds; C016300:chloramine-T; D014508:Urea; D008094:Lithium; D007784:Lactoperoxidase; D010544:Peroxidases; D018021:Lithium Chloride