Reliable semi-synthesis of hydrolysis-resistant 3'-peptidyl-tRNA conjugates containing genuine tRNA modifications.
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.
Graber
Dagmar
D
Institute of Organic Chemistry, Center for Molecular Biosciences, University of Innsbruck, 6020 Innsbruck, Austria.
Moroder
Holger
H
Steger
Jessica
J
Trappl
Krista
K
Polacek
Norbert
N
Micura
Ronald
R
D001483:Base Sequence; D021881:DNA, Catalytic / Q000378:metabolism; D006868:Hydrolysis; D008969:Molecular Sequence Data; D010455:Peptides / Q000737:chemistry*; D011130:RNA Ligase (ATP) / Q000378:metabolism; D012343:RNA, Transfer / Q000737:chemistry* / Q000378:metabolism
D021881:DNA, Catalytic; D010455:Peptides; D012343:RNA, Transfer; D011130:RNA Ligase (ATP)