Purification and Use of tRNA for Enzymatic Post-translational Addition of Amino Acids to Proteins
Irem Avcilar-Kücükgöze, Howard Gamper, Ya‐Ming Hou, Anna Kashina
Abstract
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)
Topics & Concepts
Transfer RNAAmino acidAmino Acyl-tRNA SynthetasesBiochemistryAminoacyl-tRNAEnzymeProtein biosynthesisChemistryGenetic codeTranslation (biology)BiologyComputational biologyRNAMessenger RNAGeneRNA modifications and cancerCancer-related gene regulationPeptidase Inhibition and Analysis