Studies on the Formation of Transfer Ribonucleic Acid-Ribosome Complexes

Aminoacyl transfer RNA bound to 70 S ribosomes in the presence of a template was resistant to digestion by pancreatic ribonuclease. On the basis of this observation, a convenient sensitive assay for codon recognition and aminoacyl-tRNA binding to ribosomes was described. The protection occurred as a result of the association of aminoacyl-tRNA with the 50 S subribosomal particle; thus, the assay provided a sensitive functional test involving the specific interaction of tRNA with the 50 S subunit of Escherichia coli ribosomes. The kinetics of attachment of 50 S subunits to 30 S ribosome-polyribonucleotide template-aminoacyl-tRNA complexes was studied by the assay. Furthermore, the assay provides an additional parameter for evaluating the site of action of antibiotics and other substances interfering with protein biosynthesis.