Assaying the Posttranslational Arginylation of Proteins in Cultured Cells

GALIANO M.R.; HALLAK M.E.

Protein Arginylation: Methods and Protocols, Methods in Molecular Biology

Springer Science+Business Media New York

Lugar: New York; Año: 2015;

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.