Preventing arginine-to-proline conversion in a cell-line-independent manner during cell cultivation under stable isotope labeling by amino acids in cell culture (SILAC) conditions.
Quantitative proteomics has increasingly gained impact in life science research as a tool to describe changes in protein expression between different cellular states. Stable isotope labeling by amino acids in cell culture (SILAC) is a powerful technique for relative quantification of proteins. However, the accuracy of quantification is impaired by the metabolic conversion of arginine to proline resulting in additional heavy labeled proline peptide satellites. Here we reinvestigated the addition of unlabeled proline during cell cultivation under SILAC conditions considering several thousand peptides and demonstrated that the arginine-to-proline conversion is prevented independent of the cell line used.
Functional Proteome Analysis, German Cancer Research Center, 69120 Heidelberg, Germany.
Christopher
C
Lössner
Uwe
U
Warnken
Armin
A
Pscherer
Martina
M
Schnölzer
D000596:Amino Acids / Q000737:chemistry*; D001120:Arginine / Q000378:metabolism*; D018929:Cell Culture Techniques; D003470:Culture Media / Q000378:metabolism; D057809:HEK293 Cells; D006801:Humans; D007553:Isotope Labeling / Q000379:methods*; D011392:Proline / Q000378:metabolism*; D040901:Proteomics / Q000379:methods
D000596:Amino Acids; D003470:Culture Media; D001120:Arginine; D011392:Proline