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. 1974 Oct;71(10):3951–3955. doi: 10.1073/pnas.71.10.3951

Photo-Affinity Labeling of tRNA Binding Sites in Macromolecules. I. Linking of the Phenacyl-p-azide of 4-Thiouridine in (Escherichia coli) Valyl-tRNA to 16S RNA at the Ribosomal P Site*

Ira Schwartz 1, James Ofengand 1
PMCID: PMC434304  PMID: 4610566

Abstract

The phenacyl-p-azide of 4-thiouridine in (E. coli) tRNA1Val was prepared for use as a photo-affinity probe of tRNA binding sites on ribosomes. The derivatized tRNA was 90-100% as active as control tRNA for aminoacylation, nonenzymatic binding to the ribosomal P site, elongation factor Tu(EFTu)-dependent binding to the A site, EFTu-GTP-aa-tRNA ternary complex formation, and transfer of valine into polypeptide. Irradiation of p-azidophenacyl-[3H]valyl-tRNA bound noncovalently to the ribosomal P site resulted in covalent attachment of 15-20% of the noncovalently bound tRNA to the ribosomes. The linking occurred exclusively to the 16S RNA of the 30S ribosomal subunit, thus suggesting that the region of the ribosome within 9 Å of the 4-thiouridine of tRNA, when it is bound in the P site, is solely 16S RNA.

Keywords: p-azidophenacyl bromide, tRNA function, nitrenes

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

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