QTIPS: A Novel Method of Unsupervised Determination of Isotopic Amino Acid Distribution in SILAC Experiments

Stable incorporation of labeled amino acids in cell culture is a simple approach to label proteins in vivo for mass spectrometric quantification. Full incorporation of isotopically heavy amino acids facilitates accurate quantification of proteins from different cultures, yet analysis methods for determination of incorporation are cumbersome and time-consuming. We present QTIPS, Quantification by Total Identified Peptides for SILAC, a straightforward, accurate method to determine the level of heavy amino acid incorporation throughout a population of peptides detected by mass spectrometry. Using QTIPS, we show that the incorporation of heavy amino acids in baker's yeast is unaffected by the use of prototrophic strains, indicating that auxotrophy is not a requirement for SILAC experiments in this organism. This method has general utility for multiple applications where isotopic labeling is used for quantification in mass spectrometry.

Graphical AbstractRelative abundance of isotopically distinct peptide identifications by mass spectrometry can be used for relative quantification in SILAC experiments.Figure optionsDownload high-quality image (210 K)Download as PowerPoint slide