super(18)O-labeling quantitative proteomics using an ion trap mass spectrometer

We describe a method for simultaneous identification and quantitation of proteins within complex mixtures. The method consists of super(18)O-labeling, a simple stable isotope-coding that requires merely enzymatic digestion in super(18)O-water, in combination with a capillary-liquid chromatography el...

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Bibliographic Details
Published in:Proteomics (Weinheim) 2005-01, Vol.5 (1), p.16-23
Main Authors: Sakai, Jun, Kojima, Shinich, Yanagi, Kazunori, Kanaoka, Masaharu
Format: Article
Language:English
Online Access:Get full text
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Summary:We describe a method for simultaneous identification and quantitation of proteins within complex mixtures. The method consists of super(18)O-labeling, a simple stable isotope-coding that requires merely enzymatic digestion in super(18)O-water, in combination with a capillary-liquid chromatography electrospray ion-trap mass spectrometer. In a separate experiment using the same sample and a spike test, we demonstrate that the difference ratio was calculated accurately using the super(18)O-labeling method even if the protein was part of a complex mixture. Our data also suggest that the accuracy of the quantitation can be improved by averaging the difference ratios of several peptides. In comparing our method with the isotope-coded affinity tag (ICAT) method, we show that the super(18)O-labeling method has the advantages of better recovery and fewer isotope effects. Therefore, the super(18)O-labeling method is a powerful tool for large- scale proteomics applications.
ISSN:1615-9853
DOI:10.1002/pmic.200300885