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Photo-affinity labeling of 23 S rRNA by an analog of fMet-tRNAMetf

1976; Elsevier BV; Volume: 72; Issue: 4 Linguagem: Inglês

10.1016/s0006-291x(76)80188-x

1090-2104

Nahum Sonenberg, Meir Wilchek, Ada Zamir,

Advanced biosensing and bioanalysis techniques

The technique of photo-affinity labeling has been employed to localize ribosomal components at the peptidyl-transferase center that interact with the initiator tRNA. The photo-sensitive p-azido-N-tBoc -Phe-(35S)Mot-tRNAMetf acts as an analog of fMet-tRNAMetf as it requires initiation factors and T4 mRNA for binding to 70 S ribosomes. Irradiation of the reversible binding complex results in a covalent linkage of the label to the ribosome, mostly to 23 S rRNA and specifically to its 18 S fragment. This is the first instance where an analog of the initiator tRNA has been shown to interact with 23 S rRNA, a ribosomal component likely to be involved in the interaction of donor substrates with the peptidyl transferase center.