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
Descripción
Sumario: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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