Details

Autor(en) / Beteiligte

• Finkler, Marc
• Brandt, Joshua
• Boutfol, Timothée
• Grimm, Florent
• Hartz, Philip
• Ott, Albrecht

Titel

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

Ist Teil von

• STAR protocols, 2023-09, Vol.4 (3), p.102504-102504, Article 102504

Ort / Verlag

United States: Elsevier Inc

Erscheinungsjahr

2023

Link zum Volltext

Quelle

MEDLINE

Beschreibungen/Notizen

• 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. [Display omitted] •Identification of tRNA-loaded amino acids•Characterize molecular recognition fidelity of aminoacyl-tRNA-synthetases•Mass spectrometry avoids dealing with radioactive labels Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics. 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.