Protocol to identify amino acids bound to tRNA by aminoacylation using mass spectrometry

tRNA-bound amino acids often need to be identified, for instance, in cases where different amino acids compete for binding to the same tRNA. Here, we present a mass-spectrometry-based protocol to determine the amino acids bound to tRNA by aminoacylation. We detail how to perform the aminoacylation r...

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Detalles Bibliográficos
Autores principales: Finkler, Marc, Brandt, Joshua, Boutfol, Timothée, Grimm, Florent, Hartz, Philip, Ott, Albrecht
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Elsevier 2023
Materias:
Protocol
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10436234/
https://www.ncbi.nlm.nih.gov/pubmed/37585296
http://dx.doi.org/10.1016/j.xpro.2023.102504
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author Finkler, Marc
Brandt, Joshua
Boutfol, Timothée
Grimm, Florent
Hartz, Philip
Ott, Albrecht
author_facet Finkler, Marc
Brandt, Joshua
Boutfol, Timothée
Grimm, Florent
Hartz, Philip
Ott, Albrecht
author_sort Finkler, Marc
collection PubMed
description tRNA-bound amino acids often need to be identified, for instance, in cases where different amino acids compete for binding to the same tRNA. Here, we present a mass-spectrometry-based protocol to determine the amino acids bound to tRNA by aminoacylation. We detail how to perform the aminoacylation reaction, the preparation of the aminoacyl-tRNA for measurement, and the mass spectrometric analysis. We use arginine acylation as an example; however, this protocol can be applied to any other amino acid.
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spelling pubmed-104362342023-08-19 Protocol to identify amino acids bound to tRNA by aminoacylation using mass spectrometry Finkler, Marc Brandt, Joshua Boutfol, Timothée Grimm, Florent Hartz, Philip Ott, Albrecht STAR Protoc Protocol tRNA-bound amino acids often need to be identified, for instance, in cases where different amino acids compete for binding to the same tRNA. Here, we present a mass-spectrometry-based protocol to determine the amino acids bound to tRNA by aminoacylation. We detail how to perform the aminoacylation reaction, the preparation of the aminoacyl-tRNA for measurement, and the mass spectrometric analysis. We use arginine acylation as an example; however, this protocol can be applied to any other amino acid. Elsevier 2023-08-15 /pmc/articles/PMC10436234/ /pubmed/37585296 http://dx.doi.org/10.1016/j.xpro.2023.102504 Text en © 2023 The Author(s) https://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
Finkler, Marc
Brandt, Joshua
Boutfol, Timothée
Grimm, Florent
Hartz, Philip
Ott, Albrecht
Protocol to identify amino acids bound to tRNA by aminoacylation using mass spectrometry
title Protocol to identify amino acids bound to tRNA by aminoacylation using mass spectrometry
title_full Protocol to identify amino acids bound to tRNA by aminoacylation using mass spectrometry
title_fullStr Protocol to identify amino acids bound to tRNA by aminoacylation using mass spectrometry
title_full_unstemmed Protocol to identify amino acids bound to tRNA by aminoacylation using mass spectrometry
title_short Protocol to identify amino acids bound to tRNA by aminoacylation using mass spectrometry
title_sort protocol to identify amino acids bound to trna by aminoacylation using mass spectrometry
topic Protocol
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10436234/
https://www.ncbi.nlm.nih.gov/pubmed/37585296
http://dx.doi.org/10.1016/j.xpro.2023.102504
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