Document Detail


QTIPS: a novel method of unsupervised determination of isotopic amino acid distribution in SILAC experiments.
MedLine Citation:
PMID:  20451407     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
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.
Authors:
David J Dilworth; Ramsey A Saleem; Richard S Rogers; Hamid Mirzaei; John Boyle; John D Aitchison
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Publication Detail:
Type:  Journal Article; Research Support, N.I.H., Extramural     Date:  2010-04-08
Journal Detail:
Title:  Journal of the American Society for Mass Spectrometry     Volume:  21     ISSN:  1879-1123     ISO Abbreviation:  J. Am. Soc. Mass Spectrom.     Publication Date:  2010 Aug 
Date Detail:
Created Date:  2010-08-02     Completed Date:  2010-12-03     Revised Date:  2011-09-26    
Medline Journal Info:
Nlm Unique ID:  9010412     Medline TA:  J Am Soc Mass Spectrom     Country:  United States    
Other Details:
Languages:  eng     Pagination:  1417-22     Citation Subset:  IM    
Copyright Information:
Copyright 2010 American Society for Mass Spectrometry. Published by Elsevier Inc. All rights reserved.
Affiliation:
Institute for Systems Biology, Seattle, Washington 98103, USA.
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MeSH Terms
Descriptor/Qualifier:
Amino Acid Sequence
Amino Acids / chemistry*,  metabolism
Isotope Labeling / methods*
Isotopes / chemistry*
Mass Spectrometry
Metabolic Networks and Pathways
Molecular Sequence Data
Peptide Fragments / chemistry*
Saccharomyces cerevisiae / chemistry,  metabolism
Grant Support
ID/Acronym/Agency:
GM075152/GM/NIGMS NIH HHS; GM076547/GM/NIGMS NIH HHS; P50 GM076547-04/GM/NIGMS NIH HHS; R01 GM075152-02/GM/NIGMS NIH HHS; R01 GM075152-02S1/GM/NIGMS NIH HHS; RR022220/RR/NCRR NIH HHS; U54 RR022220-05/RR/NCRR NIH HHS
Chemical
Reg. No./Substance:
0/Amino Acids; 0/Isotopes; 0/Peptide Fragments
Comments/Corrections

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine


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