Chemama Maryline, Fonvielle Matthieu, Arthur Michel, Valéry Jean-Marc, Etheve-Quelquejeu Mélanie
UMR 7613, Synthèse, Structure et Fonction de Molécules Bioactives, Université Pierre et Marie Curie, 4 place Jussieu, case 179, 75 252 Paris Cedex, France.
Chemistry. 2009;15(8):1929-38. doi: 10.1002/chem.200801563.
Aminoacyl-tRNAs have important roles in a variety of biological processes, including protein synthesis by ribosomes, targeting of proteins for degradation by the proteasome, and bacterial cell wall synthesis. Here we describe the synthesis of stable aminoacyl-tRNA analogues containing 1,4- and 1,5-substituted 1,2,3-triazole rings. The procedure involves i) Cu- and Ru-catalysed cycloadditions of 3'-azidoadenosine and alkynes, which produced the 1,4 and 1,5 regioisomers of the triazoles, respectively, ii) coupling between the resulting triazole-deoxyadenosine derivatives and a deoxycytidine phosphoramidite, and iii) the enzymatic ligation of the substituted dinucleotides with a 22 nt RNA microhelix that mimics the acceptor arm of tRNA. Nucleoside and nucleotide compounds were characterized by MS spectrometry and (1)H, (31)P and (13)C NMR spectroscopy and were assayed for inhibition of FemX(Wv), an alanyltransferase essential for the formation of the peptidoglycan network of gram-positive bacterial pathogens. The low IC(50) values obtained (2 to 4 microM) indicate that the five-membered triazole rings acted as bioisosters of esters and can be used for the design of stable aminoacyl-tRNA analogues.
氨酰基 - tRNA在多种生物过程中发挥着重要作用，包括核糖体的蛋白质合成、蛋白酶体对蛋白质的靶向降解以及细菌细胞壁合成。在此，我们描述了含有1,4 - 和1,5 - 取代的1,2,3 - 三唑环的稳定氨酰基 - tRNA类似物的合成。该过程包括：i）3'-叠氮腺苷与炔烃的铜催化和钌催化环加成反应，分别生成三唑的1,4和1,5区域异构体；ii）所得三唑 - 脱氧腺苷衍生物与脱氧胞苷亚磷酰胺之间的偶联；iii）将取代的二核苷酸与模拟tRNA受体臂的22 nt RNA微螺旋进行酶促连接。核苷和核苷酸化合物通过质谱以及（1）H、（31）P和（13）C核磁共振光谱进行表征，并检测其对FemX（Wv）的抑制作用，FemX（Wv）是革兰氏阳性细菌病原体肽聚糖网络形成所必需的丙氨酰转移酶。获得的低IC（50）值（2至4 microM）表明五元三唑环可作为酯的生物电子等排体，可用于设计稳定的氨酰基 - tRNA类似物。
