Photoincorporation of Puromycin and N-(ethyl-2-diazomalonyl)puromycin into Escherichia Coli Ribosomes

Overview

Journal Proc Natl Acad Sci U S A

Specialty Science

Date 1975 Aug 1

PMID 1103130

Citations 4

Authors

B S Cooperman

E N Jaynes

D J Brunswick

M A Luddy

Affiliations

Soon will be listed here.

Abstract

[3H]Puromycin and N-(ethyl-2-diazomalonyl)[3H]puromycin are incorporated into E. coli ribosomes on irradiation at 253.7 nm. Both compounds incorporate into both protein and nucleic acid. Two-dimensional gel electrophoresis of ribosomal protein shows that L23 is the major protein labeled by puromycin. Although incorporation is clearly a complex process, evidence is presented that L23 is labeled via an affinity labeling process, thus placing L23 at the aminoacyl-tRNA receptor (A) site. N-(ethyl-2-diazomalonyl)puromycin is a ribosomal ligand, as shown by its inhibition of two ribosomal assays, but it is not a good puromycin analog, and it is unclear whether its incorporation, which proceeds via both carbene-dependent and carbene-independent processes, results from affinity labeling.

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Identification of a region in 23S rRNA located at the peptidyl transferase center.

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