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Purification and Use of tRNA for Enzymatic Post-translational Addition of Amino Acids to Proteins

2020; Elsevier BV; Volume: 1; Issue: 3 Linguagem: Inglês

10.1016/j.xpro.2020.100207

2666-1667

Irem Avcilar-Kücükgöze, Howard Gamper, Ya‐Ming Hou, Anna Kashina,

Peptidase Inhibition and Analysis

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 tRNAArg 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)