Laboratoire de Chimie et de Biochimie Pharmacologiques et Toxicologiques, Université Paris Descartes, UMR 8601, Paris, F-75005, France.
Org Biomol Chem. 2018 Mar 14;16(11):1903-1911. doi: 10.1039/c8ob00019k.
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 A locked in South conformation. Biological evaluation of our tRNA analogues has been performed using amino-acyl tRNA-dependent transferase FemX, 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 FemX with IC of 10 and 8 μM. These results indicate that FemX displays a moderate preference for tRNAs containing a terminal A 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 tRNA product of the aminoacyl transfer reaction catalyzed by FemX.
我们在此报告了两种约束二核苷酸的合成路线，并通过对起始核苷的 NMR 分析确定了糖的构象。通过对微螺旋 RNA 的酶促连接，可获得含有 3'端 A 锁定在 South 构象的 tRNA 类似物。我们使用介导氨基酸非核糖体掺入细菌细胞壁的氨酰-tRNA 依赖性转移酶 FemX 对我们的 tRNA 类似物进行了生物学评估。我们已经表明，我们的 tRNA 类似物以 IC 为 10 和 8 μM 抑制了 FemX 催化的氨酰基转移反应。这些结果表明，FemX 对含有末端 A 锁定在 South 构象的 tRNA 表现出中等偏好，并且末端核糖的构象从 South 到 North 的转变可能有助于释放 FemX 催化的氨酰基转移反应的未带电 tRNA 产物。
