Synthesis of tRNA analogues containing a terminal ribose locked in the South conformation to study tRNA-dependent enzymes.

• 影响指数: 3.89
• 发表时间: 03 2018 14
• 来源期刊: Org Biomol Chem
• DOI: 10.1039/c8ob00019k
• PMID: 29484333
• 文章类型: Journal Article
• 作者列表: Iannazzo L,Fonvielle M,Braud E,Hřebabecký H,Procházková E,Nencka R,Mathé C,Arthur M,Etheve-Quelquejeu M

Mesh: Aminoacyltransferases / antagonists & inhibitors metabolism; Bacterial Proteins / metabolism; Chemistry Techniques, Synthetic / methods; Models, Molecular; Nucleic Acid Conformation; RNA, Transfer / chemical synthesis chemistry metabolism; Ribonucleotides / chemical synthesis chemistry metabolism; Ribose / analogs & derivatives chemical synthesis metabolism; Weissella / enzymology metabolism

Abstract

We report here the synthetic route of two constrained dinucleotides and the determination of the sugar puckering by NMR analyses of the starting nucleosides. Enzymatic ligation to microhelix-RNAs provide access to tRNA analogues containing a 3\' terminal A76 locked in South conformation. Biological evaluation of our tRNA analogues has been performed using amino-acyl tRNA-dependent transferase FemXWv, which mediates non-ribosomal incorporation of amino acids into the bacterial cell wall. We have shown that our tRNA analogues inhibited the aminoacyl transfer reaction catalyzed by FemXWv with IC50s of 10 and 8 μM. These results indicate that FemXWv displays a moderate preference for tRNAs containing a terminal A76 locked in the South conformation and that a South to North switch in the conformation of the terminal ribose might contribute to the release of the uncharged tRNAAla product of the aminoacyl transfer reaction catalyzed by FemXwv.