Abstract
The extend of the reaction between puromycin and yeast peptidyl-tRNA prelabeled in vitro was determined by measuring the distribution of trichloroacetic acid precipitable material in isokinetic sucrose gradients in the presence of 0.5 M KCl.
Thus it was found that increasing amounts of puromycin remove increasing amounts of peptidyl-tRNA from the 80S position in the gradient. The extend of the reaction, however, was independent of pretreatment of the ribosomes with inhibitors of the translocation indicating that peptidyl-tRNA at the donor and at the acceptor site of the ribosomes are equally accessible to puromycin at 0.5 M KCl.
The exposure of both ribosomal binding sites to puromycin in high salt is accompanied by an enhanced reactivity of puromycin towards peptidyl-tRNA. The ED50 determined by measuring the inhibition by puromycin of the poly-U dependent phenylalanine incorporation drops from 5×10-5 M below 250 mM KCl to 5×10-6 M at 300 mM and higher concentrations of KCl.
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Van Der Mast, C.A., Bloemers, H.P.J. The puromycin reaction mediated by yeast ribosomes in high salt. Molecular Biology Reports 1, 225–231 (1973). https://doi.org/10.1007/BF00357646
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DOI: https://doi.org/10.1007/BF00357646