Assaying the Posttranslational Arginylation of Proteins in Cultured Cells.
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 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.
Departamento de Química Biológica, Facultad de Ciencias Químicas, Centro de Investigaciones en Química Biológica de Córdoba, CIQUIBIC, (UNC-CONICET), Universidad Nacional de Córdoba (X5000HUA), Córdoba, Argentina.
Mauricio R
MR
Galiano
Marta E
ME
Hallak
D019881:Aminoacyltransferases / Q000378:metabolism; D000818:Animals; D001120:Arginine / Q000378:metabolism*; D002460:Cell Line; D002478:Cells, Cultured; D007553:Isotope Labeling; D011499:Protein Processing, Post-Translational; D011506:Proteins / Q000737:chemistry / Q000378:metabolism*; D020539:Sequence Analysis, Protein / Q000379:methods; D013194:Staining and Labeling
D011506:Proteins; D001120:Arginine; D019881:Aminoacyltransferases