Photoaffinity Labeling of the Ribosomal Peptidyl Transferase Site with Synthetic Puromycin Analogues

Robert R. Vince, Jay Brownell, Keilailau L. Fong

Research output: Contribution to journalArticlepeer-review

8 Scopus citations

Abstract

A photoaffinity labeling puromycin analogue, N-(2-nitro-4-azidophenyl)-L-lysinyl puromycin aminonucleoside (NAP-Lys-Pan), was synthesized and used for investigation of the peptidyl transferase center of 70S ribosomes. Visible light irradiation of NAP-Lys-Pan led to covalent linkage of the analogue with Escherichia coli ribosomes. In a subsequent step, poly(uridylic acid) was employed to direct Ac[14C]Phe-tRNA to the P sites of the photolabeled ribosomes. Transpeptidation of Ac[14C]phenylalanine to the bound NAP-Lys-Pan resulted in selective incorporation of radioactive label into the peptidyl transferase A site. Dissociation of the ribosomes into subunits, and digestion of the RNA components, indicated that the radioactive label was incorporated into a protein fraction of the 50S subunit.

Original languageEnglish (US)
Pages (from-to)5489-5493
Number of pages5
JournalBiochemistry
Volume17
Issue number25
DOIs
StatePublished - Jan 1 1978

Fingerprint Dive into the research topics of 'Photoaffinity Labeling of the Ribosomal Peptidyl Transferase Site with Synthetic Puromycin Analogues'. Together they form a unique fingerprint.

Cite this