Modified aminoacyl-tRNA. III. A general procedure for the synthesis of dipeptidyl transfer RNA

Y. Lapidot*, N. de Groot, S. Rappoport, A. D. Hamburger

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

17 Scopus citations

Abstract

A general method for the preparation of dipeptidyl transport RNA (tRNA) is described. The synthesis involves a reaction between N-hydroxysuccinimide esters of N-monomethoxytritylamino acids and aminoacyl-tRNA. The activated N-protected amino acids react specifically with the amino group of the amino acid attached to the tRNA and do not react with any of the functional groups of the tRNA. [14C]Valyl-, [14C]phenylalanyl- and [14C]methionyl-tRNA could be 100 % transformed into the corresponding dipeptidyl-tRNA when they were reacted with activated, N-protected glycine and alanine. With the activated N-protected phenylalanine only 80-85 % of dipeptidyl-tRNA was formed. Mono-p-methoxytrityl, notwithstanding its bulkiness12, was found suitable for protecting the amino group of several amino acids. In all the reactions tested, the monomethoxytrityl remained attached to the amino group during the acylation and could be removed from the dipeptidyl-tRNA under mild acidic conditions. The tRNA of the dipeptidyl-tRNA did not lose its biological properties as a result of the chemical synthesis. Dipeptidyl-tRNA could be bound to ribosomes in a polyuridylic acid dependent reaction, and the tRNA could also be recharged with amino acid after removal of the dipeptide from the tRNA by mild alkaline hydrolysis.

Original languageEnglish
Pages (from-to)532-539
Number of pages8
JournalBBA Section Nucleic Acids And Protein Synthesis
Volume149
Issue number2
DOIs
StatePublished - 19 Dec 1967

Fingerprint

Dive into the research topics of 'Modified aminoacyl-tRNA. III. A general procedure for the synthesis of dipeptidyl transfer RNA'. Together they form a unique fingerprint.

Cite this