Physiol. Genomics Ad Instruments
HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH
QUICK SEARCH:   [advanced]


     

Physiol. Genomics (August 7, 2007). doi:10.1152/physiolgenomics.00096.2007
This Article
Right arrow Full Text (PDF)
Right arrow All Versions of this Article:
31/2/357    most recent
00096.2007v1
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Services
Right arrow Email this article to a friend
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Download to citation manager
Citing Articles
Right arrow Citing Articles via HighWire
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Smith, J.
Right arrow Articles by Greene, A. S
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Smith, J.
Right arrow Articles by Greene, A. S
Submitted on April 27, 2007
Accepted on August 5, 2007

Relative quantification of peptide phosphorylation in a complex mixture using 18O labeling

Julia Smith1, Michael Olivier2, and Andrew S Greene3*

1 Biotechnology and Bioengineering Center, Medical College of Wisconsin, Milwaukee, Wisconsin, United States
2 Biotechnology and Bioengineering Center, Medical College of Wisconsin, Milwaukee, Wisconsin, United States; Human & Molecular Genetics Center, Dept. of Physiology, Medical College of Wisconsin, Milwaukee, Wisconsin, United States
3 Biotechnology and Bioengineering Center, Medical College of Wisconsin, Milwaukee, Wisconsin, United States; Physiology, Medical College of Wisconsin, Milwaukee, Wisconsin, United States

* To whom correspondence should be addressed. E-mail: agreene{at}mcw.edu.

We have developed a method to determine the degree of phosphorylation of a peptide in a complex mixture without enrichment or operation of the mass spectrometer in negative ion mode. Yeast lysate containing known amounts of synthetic peptides (VPQLEIVPNSAEERLHSMK and VPQLEIVPN[pS]AEERLHSMK) was labeled with 16O and 18O during hydrolysis. After treatment of one sample with a cocktail of phosphatases the two samples were pooled. The intensity of the dephosphorylated peptide peaks was used to infer the degree of phosphorylation present before treatment. The linear dynamic range of this method is greater than ten fold before either of the peptide envelopes becomes indistinguishable from the surrounding noise. Since both the site of post translational modification, and the proportion of the protein population that is modified is vital in protein function, the employment of this technique will provide a valuable tool for the analysis of the functional implications of protein phosphorylation.




This article has been cited by other articles:


Home page
 Mol. Cell. ProteomicsHome page
K. A. Dave, F. Whelan, C. Bindloss, S. G. B. Furness, A. Chapman-Smith, M. L. Whitelaw, and J. J. Gorman
Sulfonation and Phosphorylation of Regions of the Dioxin Receptor Susceptible to Methionine Modifications
Mol. Cell. Proteomics, April 1, 2009; 8(4): 706 - 719.
[Abstract] [Full Text] [PDF]

Home page
 Brief Funct Genomic ProteomicHome page
X. Ye, B. Luke, T. Andresson, and J. Blonder
18O Stable Isotope Labeling in MS-based Proteomics
Brief Funct Genomic Proteomic, March 1, 2009; 8(2): 136 - 144.
[Abstract] [Full Text] [PDF]

Home page
 Physiol. GenomicsHome page
S. P. Mirza and M. Olivier
Methods and approaches for the comprehensive characterization and quantification of cellular proteomes using mass spectrometry
Physiol Genomics, October 8, 2008; 33(1): 3 - 11.
[Abstract] [Full Text] [PDF]


HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH
Visit Other APS Journals Online
Copyright © 2007 by the American Physiological Society.