A detergent-free method for isolation of viable ribosomes from rat brain.
A detergent-free method based on sucrose density gradient centrifugations was developed for isolation of free ribosomes from rat brain. The purity of the preparations was checked by RNA/protein ratio, electron microscopy and marker enzymes. They were uncontaminated by other cell organelles. The incorporation of leucine, isoleucine and valine into acid-insoluble fraction by ribosomes comformed to the Hill equation v/V = sn/(K+sn). Branched-chain amino acids mutually inhibited each other's incorporation by ribosomes, but the inhibition was abolished with detergents frequently used in isolation procedures.
D000596:Amino Acids / Q000378:metabolism; D000818:Animals; D001921:Brain / Q000648:ultrastructure*; D002458:Cell Fractionation; D009419:Nerve Tissue Proteins / Q000378:metabolism; D012313:RNA / Q000378:metabolism; D051381:Rats; D012260:Ribonucleases / Q000037:antagonists & inhibitors; D012270:Ribosomes / Q000032:analysis* / Q000201:enzymology
D000596:Amino Acids; D009419:Nerve Tissue Proteins; D012313:RNA; D012260:Ribonucleases