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

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...

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Detalles Bibliográficos
Autores principales: Avcilar-Kucukgoze, Irem, Gamper, Howard, Hou, Ya-Ming, Kashina, Anna
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Elsevier 2020
Materias:
Protocol
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7757669/
https://www.ncbi.nlm.nih.gov/pubmed/33377101
http://dx.doi.org/10.1016/j.xpro.2020.100207
Descripción
Sumario: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(Arg) 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)

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