A Genetic Engineering Solution to the Arginine Conversion Problem in Stable Isotope Labeling by Amino Acids in Cell Culture (SILAC)
Sawin lab paper featured in Molecular & Cellular Proteomics.
Authors
Bicho C., de Lima F. Chen Z, Rappsilber J and Sawin KE
Summary
The Sawin/Rappsilber labs have developed a new strategy to avoid a problem associated with SILAC analysis: the in vivo conversion of labeled arginine to other amino acids. Bicho and coauthors avoided this problem using a simple but efficient genetic engineering trick (the deletion of genes involved in the catabolism of arginine) and a slight modification of the growing medium of the yeast mutant strains. The figure shows how this new fission yeast SILAC methodology can be applied successfully to the identification of both proteins that were up-regulated and proteins that were down-regulated as cells progressed from G2 to G1/S.