[45] Isolation of mammalian ribosomal subunits active in polypeptide synthesis

摘要: Publisher Summary This chapter aims to develop a method for the preparation and purification of active ribosomal subunits from rat mouse liver. The technique involves incubating purified polysomes with all components protein synthesis until competent ribosomes terminate release their polypeptide chain. Upon treatment such an incubation mixture, are found as 60 S 40 subunits. subunits, when recombined, spontaneously associate 80 couples in absence tRNA, mRNA, or supernatant factors. subunit contains enzymatic site catalysis peptide bond formation. Purified liver able catalyze formation bonds using substrates puromycin N-formyl- N-acetylaminoacyl oligonucleotides tRNA. A relative comparison rates N-acetylleucyl-puromycin catalyzed by E. coli 50 shows that reaction rate is approximately twice