Abstract
This chapter describes the preparation of pre-charged Arg-tRNA that can be used in arginylation reaction. While in a typical arginylation reaction arginyl-tRNA synthetase (RARS) is normally included as a component of the reaction and continually charges tRNA during arginylation, it is sometimes necessary to separate the charging and the arginylation step, in order to perform each reaction under controlled conditions, e.g., for measuring the kinetics or determining the effect of different compounds and chemicals on the reaction. In such cases, tRNAArg can be pre-charged with Arg and purified away from the RARS enzyme prior to arginylation.
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References
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Acknowledgments
This work was supported by NIH R35GM122505 and R01NS102435 to A.K.
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Avcilar-Kucukgoze, I., Kashina, A.S. (2023). Enzymatic Aminoacylation of tRNAArg Using Recombinant Arg-tRNA Synthetase. In: Kashina, A.S. (eds) Protein Arginylation. Methods in Molecular Biology, vol 2620. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-2942-0_14
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DOI: https://doi.org/10.1007/978-1-0716-2942-0_14
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