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.
Dilworth
David J
DJ
Institute for Systems Biology, Seattle, Washington 98103, USA.
Saleem
Ramsey A
RA
Rogers
Richard S
RS
Mirzaei
Hamid
H
Boyle
John
J
Aitchison
John D
JD
D000595:Amino Acid Sequence; D000596:Amino Acids / Q000737:chemistry* / Q000378:metabolism; D007553:Isotope Labeling / Q000379:methods*; D007554:Isotopes / Q000737:chemistry*; D013058:Mass Spectrometry; D053858:Metabolic Networks and Pathways; D008969:Molecular Sequence Data; D010446:Peptide Fragments / Q000737:chemistry*; D012441:Saccharomyces cerevisiae / Q000737:chemistry / Q000378:metabolism
D000596:Amino Acids; D007554:Isotopes; D010446:Peptide Fragments
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