A chemical mapping technique for exploring the location of proteins along the ribosome-bound peptide chain

Dan Eilat*, Maria Pellegrini, Helen Oen, Yehuda Lapidot, Charles R. Cantor

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

21 Scopus citations

Abstract

A series of peptidyl-tRNA analogs with varying peptide chain length, BrAc(Gly) nPhe-tRNAphe, n = 0 to 16 has been prepared. When bound to Escherichia coli 70 S ribosomes these all react covalently with certain ribosomal proteins. The overwhelming majority of the reaction is with 50 S ribosomal proteins L2, L16, L24, L26-L27 and L32-L33. The extent of reaction with each protein is a function of peptide chain length, making it possible to estimate the relative proximity of these proteins to the 3′-terminus of tRNA bound in the ribosomal P site. This fact, coupled with the findings of others about the length dependence of the binding and peptide donor activity of peptidyl-tRNAs suggests that there is actually a binding site for the growing peptide chain. If this is true, the results presented here permit the ordering of the proteins in this site: L2 is closest to the 3′-end of tRNA followed by L26-L27, L32-L33 and last L24. Evidence is also given that the direction of the growing peptide chain must point away from the A site.

Original languageEnglish
Pages (from-to)831-840
Number of pages10
JournalJournal of Molecular Biology
Volume88
Issue number4
DOIs
StatePublished - 5 Oct 1974

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