HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS		QUICK SEARCH:   [advanced] Author: Keyword(s): Year:  Vol:  Page:

This Article Full Text Full Text (PDF) All Versions of this Article: M300001-MCP200v1 2/3/138    most recent Submit a response Alert me when this article is cited Alert me when eLetters are posted Alert me if a correction is posted Citation Map Services Email this article to a friend Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Glossary Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Steen, H. Articles by Mann, M. Search for Related Content PubMed PubMed Citation Articles by Steen, H. Articles by Mann, M. Social Bookmarking                      What's this?

Molecular & Cellular Proteomics 2:138-145, 2003.
© 2003 by The American Society for Biochemistry and Molecular Biology, Inc.

---

Research

Phosphotyrosine Mapping in Bcr/Abl Oncoprotein Using Phosphotyrosine-specific Immonium Ion Scanning^*

Hanno Steen, Minerva Fernandez, Saghi Ghaffari^,¶, Akhilesh Pandey^|| and Matthias Mann^**

Center for Experimental Bioinformatics, Department of Biochemistry and Molecular Biology, University of Southern Denmark, 5230 Odense M, Denmark
Whitehead Institute for Biomedical Research and Massachusetts Institute of Technology, Cambridge, Massachusetts 02142

Bcr/Abl is a fusion oncoprotein that is of paramount importance in chronic myelogenous leukemia and acute lymphocytic leukemia. The tyrosine-phosphorylated fraction of the p185 form of Bcr/Abl was isolated by immunoprecipitation with an anti-phosphotyrosine antibody and SDS-PAGE. The tryptic digest of the gel-separated protein was prefractionated on POROS R2/OLIGO R3 microcolumns and subjected to phosphotyrosine mapping by precursor ion scanning in positive ion mode utilizing the immonium ion of phosphotyrosine, also called phosphotyrosine-specific immonium ion scanning, on a quadrupole time-of-flight tandem mass spectrometer. In total, nine different phosphorylated tyrosine residues were unambiguously localized in 12 different precursor ions. These phosphorylation sites correspond to three previously described phosphotyrosine residues and six novel tyrosine phosphorylation sites, and most of them were not predicted by the phosphorylation motif prediction programs ProSite, NetPhos, or ScanSite. This study shows the power of phosphotyrosine-specific immonium ion scanning for sensitive phosphotyrosine mapping when limited amounts of samples are available.

^** To whom correspondence may be addressed: Center for Experimental Bioinformatics, Dept. of Biochemistry and Molecular Biology, University of Southern Denmark, Campusvej 55, 5230 Odense M, Denmark. E-mail: mann{at}bmb.sdu.dk

CiteULike    Complore    Connotea    Del.icio.us    Digg    Reddit    Technorati    What's this?

This article has been cited by other articles:

				V. L. Goss, K. A. Lee, A. Moritz, J. Nardone, E. J. Spek, J. MacNeill, J. Rush, M. J. Comb, and R. D. Polakiewicz A common phosphotyrosine signature for the Bcr-Abl kinase Blood, June 15, 2006; 107(12): 4888 - 4897. [Abstract] [Full Text] [PDF]

				E. Boeri Erba, E. Bergatto, S. Cabodi, L. Silengo, G. Tarone, P. Defilippi, and O. N. Jensen Systematic Analysis of the Epidermal Growth Factor Receptor by Mass Spectrometry Reveals Stimulation-dependent Multisite Phosphorylation Mol. Cell. Proteomics, August 1, 2005; 4(8): 1107 - 1121. [Abstract] [Full Text] [PDF]

				I. M. Cristea, S. J. Gaskell, and A. D. Whetton Proteomics techniques and their application to hematology Blood, May 15, 2004; 103(10): 3624 - 3634. [Abstract] [Full Text] [PDF]