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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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
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author Avcilar-Kucukgoze, Irem
Gamper, Howard
Hou, Ya-Ming
Kashina, Anna
author_facet Avcilar-Kucukgoze, Irem
Gamper, Howard
Hou, Ya-Ming
Kashina, Anna
author_sort Avcilar-Kucukgoze, Irem
collection PubMed
description 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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spelling pubmed-77576692020-12-28 Purification and Use of tRNA for Enzymatic Post-translational Addition of Amino Acids to Proteins Avcilar-Kucukgoze, Irem Gamper, Howard Hou, Ya-Ming Kashina, Anna STAR Protoc Protocol 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) Elsevier 2020-12-09 /pmc/articles/PMC7757669/ /pubmed/33377101 http://dx.doi.org/10.1016/j.xpro.2020.100207 Text en © 2020 The Authors http://creativecommons.org/licenses/by-nc-nd/4.0/ This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).
spellingShingle Protocol
Avcilar-Kucukgoze, Irem
Gamper, Howard
Hou, Ya-Ming
Kashina, Anna
Purification and Use of tRNA for Enzymatic Post-translational Addition of Amino Acids to Proteins
title Purification and Use of tRNA for Enzymatic Post-translational Addition of Amino Acids to Proteins
title_full Purification and Use of tRNA for Enzymatic Post-translational Addition of Amino Acids to Proteins
title_fullStr Purification and Use of tRNA for Enzymatic Post-translational Addition of Amino Acids to Proteins
title_full_unstemmed Purification and Use of tRNA for Enzymatic Post-translational Addition of Amino Acids to Proteins
title_short Purification and Use of tRNA for Enzymatic Post-translational Addition of Amino Acids to Proteins
title_sort purification and use of trna for enzymatic post-translational addition of amino acids to proteins
topic Protocol
url 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
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