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Detection of Protein Ubiquitination Sites by Peptide Enrichment and Mass Spectrometry
Published on: March 23, 2020
Assaying the Posttranslational Arginylation of Proteins in Cultured Cells
Mauricio R Galiano1, Marta E Hallak2
1Dpt. Quimica Biologica Ranwel Caputto, Facultad de Ciencias Quimicas, Universidad Nacional de Córdoba, CIQUIBIC-CONICET, Córdoba, Argentina.
Abstract:
To evaluate the posttranslational arginylation of proteins in vivo, we describe a protocol for studying the 14C-Arg incorporation into proteins of cells in culture. The conditions determined for this particular modification contemplate both the biochemical requirements of the enzyme ATE1 and the adjustments that allowed the discrimination between posttranslational arginylation of proteins and de novo synthesis. These conditions are applicable for different cell lines or primary cultures, representing an optimal procedure for the identification and the validation of putative ATE1 substrates.

