Synthesis of stable aminoacyl-tRNA analogs

• Published in: Current protocols in nucleic acid chemistry Vol. Chapter 4; p. Unit 4.44
• Main Authors: Chemama, Maryline, Fonvielle, Matthieu, Lecerf, Maxime, Mellal, Dénia, Fief, Hélène, Arthur, Michel, Etheve-Quelquejeu, Mélanie
• Format: Journal Article
• Language: English
• Published: United States 01.03.2011
• Subjects: Anti-Bacterial Agents - chemical synthesis; RNA, Transfer, Amino Acyl - chemical synthesis; RNA, Transfer, Amino Acyl - chemistry; Spectrum Analysis; Structure-Activity Relationship; Transferases - antagonists & inhibitors; Triazoles - chemistry
• DOI: 10.1002/0471142700.nc0444s44

Aminoacyl-tRNAs have important roles in a variety of biological processes. Here, we describe the synthesis of stable aminoacyl-tRNA analogs containing 1,4-substituted 1,2,3-triazole rings. The procedure involves (i) copper-catalyzed cycloadditions of 3'-or 2'-azido-adenosine and alkynes, (ii) coupling between the resulting triazole-deoxyadenosine derivatives and a deoxycytidine phosphoramidite, and (iii) the enzymatic ligation of the 2'- or 3'-triazole-dinucleotides with a 22-nt RNA microhelix that mimics the acceptor arm of tRNA. Each nucleoside and nucleotide intermediate was characterized by MS spectrometry and (1)H, (31)P, and (13)C NMR spectroscopy, and the tRNA-analogs were assayed for inhibition of FemXWv, an alanyl-transferase essential for the formation of the peptidoglycan network of Gram-positive bacterial pathogens. The low IC(50) values obtained (2 to 4 µM) indicate that the five-membered triazole rings acted as an isosteres of esters and can be used for the design of stable aminoacyl-tRNA analogs.