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J Am Soc Mass Spectrom. 2010 Aug;21(8):1417-22. doi: 10.1016/j.jasms.2010.04.002. Epub 2010 Apr 8.

QTIPS: a novel method of unsupervised determination of isotopic amino acid distribution in SILAC experiments.

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  • 1Institute for Systems Biology, Seattle, Washington 98103, USA.

Abstract

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.

Copyright 2010 American Society for Mass Spectrometry. Published by Elsevier Inc. All rights reserved.

PMID:
20451407
[PubMed - indexed for MEDLINE]
PMCID:
PMC2914207
Free PMC Article

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