使用笼化氨酰 - tRNA的光依赖性蛋白质生物合成。

Photo-dependent protein biosynthesis using a caged aminoacyl-tRNA.

作者信息

Akahoshi Akiya, Doi Yoshio, Sisido Masahiko, Watanabe Kazunori, Ohtsuki Takashi

出版信息

Bioorg Med Chem Lett. 2014 Dec 1;24(23):5369-72. doi: 10.1016/j.bmcl.2014.10.053.

DOI:10.1016/j.bmcl.2014.10.053

Abstract

Translation systems with four-base codons provide a powerful strategy for protein engineering and protein studies because they enable site-specific incorporation of non-natural amino acids into proteins. In this study, a caged aminoacyl-tRNA with a four-base anticodon was synthesized. The caged aminoacyl-tRNA contains a photocleavable nitroveratryloxycarbonyl (NVOC) group. This study showed that the caged aminoacyl-tRNA was not deacylated, did not bind to EF-Tu, and was activated by light. Photo-dependent translation of an mRNA containing the four-base codon was demonstrated using the caged aminoacyl-tRNA.

摘要

具有四碱基密码子的翻译系统为蛋白质工程和蛋白质研究提供了一种强大的策略，因为它们能够将非天然氨基酸位点特异性地掺入蛋白质中。在本研究中，合成了一种带有四碱基反密码子的笼化氨酰-tRNA。该笼化氨酰-tRNA含有一个可光裂解的硝基藜芦氧基羰基（NVOC）基团。本研究表明，该笼化氨酰-tRNA不会脱酰基，不与EF-Tu结合，且可被光激活。使用该笼化氨酰-tRNA证明了含四碱基密码子的mRNA的光依赖性翻译。