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).
Avcilar-Kucukgoze, Irem; Gamper, Howard; Hou, Ya-Ming; and Kashina, Anna, "Purification and Use of tRNA for Enzymatic Post-translational Addition of Amino Acids to Proteins." (2020). Department of Biochemistry and Molecular Biology Faculty Papers. Paper 187.
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