tRNA 的纯化及在酶促的蛋白质翻译后氨基酸添加中的应用。

Department of Biomedical Sciences, School of Veterinary Medicine, University of Pennsylvania, Philadelphia, PA 19104, USA.

Department of Biochemistry and Molecular Biology, Thomas Jefferson University, Philadelphia, PA 19144, USA.

STAR Protoc. 2020 Dec 9;1(3):100207. doi: 10.1016/j.xpro.2020.100207. eCollection 2020 Dec 18.

Post-translational addition of amino acids to proteins by enzymes using aminoacyl-tRNA is an emerging regulatory mechanism. Examples include Arg transfer in eukaryotes, Leu/Phe transfer in bacteria, and tRNA-synthetase-mediated addition of amino acids to Lys side chains. Here, we present a method of purification and use of tRNA for such reactions, focusing on tRNA and its use for arginylation. This method can also be used for other tRNA-mediated reactions. For complete details on the use and execution of this protocol, please refer to Avcilar-Kucukgoze et al. (2020).

酶利用氨酰-tRNA 将氨基酸添加到蛋白质中的翻译后修饰是一种新兴的调控机制。例如，真核生物中的 Arg 转移、细菌中的 Leu/Phe 转移以及 tRNA 合成酶介导的赖氨酸侧链氨基酸添加。在这里，我们介绍了一种用于此类反应的 tRNA 纯化和使用方法，重点介绍 tRNA 及其在精氨酸化中的应用。该方法也可用于其他 tRNA 介导的反应。有关此方案使用和执行的完整详细信息，请参阅 Avcilar-Kucukgoze 等人。(2020)。