Chemically Acylated tRNAs are Functional in Zebrafish Embryos.
Chemically Acylated tRNAs are Functional in Zebrafish Embryos.
复制标题
DOI：
10.1021/jacs.2c11452
复制
发表时间：
2023-01
影响因子：
15
通讯作者：
Wes Brown;J. Galpin;Carolyn Rosenblum;M. Tsang;C. Ahern;A. Deiters
中科院分区：
文献类型：
--
作者：
Wes Brown;J. Galpin;Carolyn Rosenblum;M. Tsang;C. Ahern;A. Deiters
遗传密码的扩展已经推动蛋白质化学超越了经典的22种氨基酸。使这成为可能的关键酶是工程氨酰tRNA合成酶。然而，随着多年来遗传编码的氨基酸数量的增加，合成酶可容纳的新型侧链的类型和大小的明显限制变得明显。在这里，我们表明，化学酰化的tRNA允许强大的，位点特异性的非天然氨基酸掺入到斑马鱼胚胎的蛋白质中，斑马鱼胚胎是人类健康和发育的重要模式生物。我们采用这种方法，将一个独特的photocaged组氨酸类似物合成酶工程的努力已经失败。此外，我们证明了光学控制不同的酶在活胚胎中安装photocaged组氨酸到其活性位点。
Genetic code expansion has pushed protein chemistry past the canonical 22 amino acids. The key enzymes that make this possible are engineered aminoacyl tRNA synthetases. However, as the number of genetically encoded amino acids has increased over the years, obvious limits in the type and size of novel side chains that can be accommodated by the synthetase enzyme become apparent. Here, we show that chemically acylating tRNAs allow for robust, site-specific incorporation of unnatural amino acids into proteins in zebrafish embryos, an important model organism for human health and development. We apply this approach to incorporate a unique photocaged histidine analogue for which synthetase engineering efforts have failed. Additionally, we demonstrate optical control over different enzymes in live embryos by installing photocaged histidine into their active sites.