Studies on Protein Synthesis in Vitro

The release of radioactive amino acid from the particulate fractions separated from the prelabelled livers of rats by centrifugation has been studied under various conditions. Although pure fractions may not have been obtained, great differences in behavior were observed. In the mitochondria and nuclei dinitrophenol (10(-4)M) causes an inhibition of release, but in microsomes the opposite effect is observed. When the incubation medium is fortified with ATP and phosphocreatine, release is inhibited. In microsomes and nuclei the inhibition proceeds to the extent that the incorporation of preformed radioactive amino acid occurs. Protein is synthesized at a rapid rate. In incubations longer than 1 hour there is always a release of radioactive amino acid. It is concluded from these results that the interpretation of release data from slices or systems such as those studied is impossible without further information concerning some of the unknown variables. The most important unknown is the specific activity of the "free" amino acid in the particulates and the effect of carrier amino acid in the medium of this specific activity.