Assaying the Posttranslational Arginylation of Proteins in Cultured Cells

MAURICIO R. GALIANO; MARTA E. HALLAK

Protein Arginylation Methods and Protocols

Methods in Molecular Biology - Human Press

Lugar: New York; Año: 2015; p. 49 - 58

To evaluate the posttranslational arginylation of proteins in vivo, we describe a protocol for studying the 14 C-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 adjustmentsthat 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.