Reliable semi-synthesis of hydrolysis-resistant 3'-peptidyl-tRNA conjugates containing genuine tRNA modifications

Nucleic Acids Res. 2010 Oct;38(19):6796-802. doi: 10.1093/nar/gkq508. Epub 2010 Jun 4.

Abstract

The 3'-peptidyl-tRNA conjugates that possess a hydrolysis-resistant ribose-3'-amide linkage instead of the natural ester linkage would represent valuable substrates for ribosomal studies. Up to date, access to these derivatives is severely limited. Here, we present a novel approach for the reliable synthesis of non-hydrolyzable 3'-peptidyl-tRNAs that contain all the respective genuine nucleoside modifications. In short, the approach is based on tRNAs from natural sources that are site-specifically cleaved within the TΨC loop by using DNA enzymes to obtain defined tRNA 5'-fragments carrying the modifications. After dephosphorylation of the 2',3'-cyclophosphate moieties from these fragments, they are ligated to the respective 3'-peptidylamino-tRNA termini that were prepared following the lines of a recently reported solid-phase synthesis. By this novel concept, non-hydrolyzable 3'-peptidyl-tRNA conjugates possessing all natural nucleoside modifications are accessible in highly efficient manner.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Base Sequence
  • DNA, Catalytic / metabolism
  • Hydrolysis
  • Molecular Sequence Data
  • Peptides / chemistry*
  • RNA Ligase (ATP) / metabolism
  • RNA, Transfer / chemistry*
  • RNA, Transfer / metabolism

Substances

  • DNA, Catalytic
  • Peptides
  • RNA, Transfer
  • RNA Ligase (ATP)