Chemical inactivation of Escherichia coli 30-S ribosomes by iodination. Identification of proteins involved in tRNA binding.
30-S ribosomal subunits are inactivated by iodination for both enzymic fMet-tRNA and non-enzymic Phe-tRNA binding activities. This inactivation is due to modification of the protein moiety of the ribosome. Reconstitutions were performed with 16-S RNA and mixtures of total protein isolated from modified subunits and purified proteins isolated from unmodified subunits. This allowed identification of the individual proteins which restore tRNA binding activity. S3, S14 and S19 were identified as proteins involved in fMet-tRNA binding. S1, S2, S3, S14 and S19 were identified as proteins involved in Phe-tRNA binding. Modified particles shown normal sedimentation constants and complete protein compositions both before and after reconstitution. This suggests that the loss of activity is due to modification of one or more of the actual binding sites located on the 30-S subunit and that restoration of activity is due to structural correction at this site rather than to correction of an assembly defect.
D001665:Binding Sites; D004926:Escherichia coli / Q000187:drug effects / Q000378:metabolism*; D007455:Iodine / Q000494:pharmacology*; D007467:Iodoproteins / Q000378:metabolism*; D008715:Methionine; D010649:Phenylalanine; D011485:Protein Binding; D012335:RNA, Ribosomal / Q000378:metabolism; D012343:RNA, Transfer / Q000378:metabolism*; D012269:Ribosomal Proteins / Q000378:metabolism*; D012270:Ribosomes / Q000187:drug effects / Q000378:metabolism*
D007467:Iodoproteins; D012335:RNA, Ribosomal; D012269:Ribosomal Proteins; D010649:Phenylalanine; D012343:RNA, Transfer; D007455:Iodine; D008715:Methionine