Advertisement
MCP
HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS
QUICK SEARCH:   [advanced]


     

Originally published In Press as doi:10.1074/mcp.M500210-MCP200 on October 31, 2005.
This Article
Right arrow Full Text
Right arrow Full Text (PDF)
Right arrow All Versions of this Article:
M500210-MCP200v1
5/2/337    most recent
Right arrow Submit a response
Right arrow Alert me when this article is cited
Right arrow Alert me when eLetters are posted
Right arrow Alert me if a correction is posted
Right arrow Citation Map
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
Right arrowRequest Permissions
Right arrow Glossary
Citing Articles
Right arrow Citing Articles via HighWire
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Williamson, B. L.
Right arrow Articles by Morrice, N. A.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Williamson, B. L.
Right arrow Articles by Morrice, N. A.
Social Bookmarking
Add to CiteULike   Add to Complore   Add to Connotea   Add to Del.icio.us   Add to Digg   Add to Reddit   Add to Technorati  
What's this?

Molecular & Cellular Proteomics 5:337-346, 2006.
© 2006 by The American Society for Biochemistry and Molecular Biology, Inc.

Research

Automated Identification and Quantification of Protein Phosphorylation Sites by LC/MS on a Hybrid Triple Quadrupole Linear Ion Trap Mass Spectrometer*

Brian L. Williamson{ddagger}, Jason Marchese{ddagger} and Nicholas A. Morrice§,¶

From the {ddagger} Discovery Proteomics Small Molecule Research Center, Applied Biosystems, Framingham, Massachusetts 01701 and the § Medical Research Council Protein Phosphorylation Unit, School of Life Sciences, University of Dundee, Dundee DD1 5EH, Scotland, United Kingdom

Complete phosphorylation mapping of protein kinases was successfully undertaken using an automated LC/MS/MS approach. This method uses the direct combination of triple quadrupole and ion trapping capabilities in a hybrid triple quadrupole linear ion trap to selectively identify and sequence phosphorylated peptides. In particular, the use of a precursor ion scan of m/z –79 in negative ion mode followed by an ion trap high resolution scan (an enhanced resolution scan) and a high sensitivity MS/MS scan (enhanced product ion scan) in positive mode is a very effective method for identifying phosphorylation sites in proteins at low femtomole levels. Coupling of this methodology with a stable isotope N-terminal labeling strategy using iTRAQTM reagents enabled phosphorylation mapping and relative protein phosphorylation levels to be determined between the active and inactive forms of the protein kinase MAPKAPK-1 in the same LC/MS run.

Funded by the Medical Research Council, UK and by Pfizer, Astra Zeneca, GlaxoSmithKline, Boehringer Ingelheim, Merck & Co., and Merck KGaA. To whom correspondence should be addressed: Medical Research Council Protein Phosphorylation Unit, School of Life Sciences, University of Dundee, Dow St., Dundee DD1 5EH, Scotland, UK. Tel.: 44-1382-348299; Fax: 44-1382-223778; E-mail: n.a.morrice{at}dundee.ac.uk


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


This article has been cited by other articles:


Home page
 Cancer Res.Home page
S. C. Brady, M. L. Coleman, J. Munro, S. M. Feller, N. A. Morrice, and M. F. Olson
Sprouty2 Association with B-Raf Is Regulated by Phosphorylation and Kinase Conformation
Cancer Res., September 1, 2009; 69(17): 6773 - 6781.
[Abstract] [Full Text] [PDF]

Home page
 J. Virol.Home page
T. W. Wisner, C. C. Wright, A. Kato, Y. Kawaguchi, F. Mou, J. D. Baines, R. J. Roller, and D. C. Johnson
Herpesvirus gB-Induced Fusion between the Virion Envelope and Outer Nuclear Membrane during Virus Egress Is Regulated by the Viral US3 Kinase
J. Virol., April 1, 2009; 83(7): 3115 - 3126.
[Abstract] [Full Text] [PDF]

Home page
 Proc. Natl. Acad. Sci. USAHome page
H. Smith, M. Peggie, D. G. Campbell, F. Vandermoere, E. Carrick, and P. Cohen
Inaugural Article: Identification of the phosphorylation sites on the E3 ubiquitin ligase Pellino that are critical for activation by IRAK1 and IRAK4
PNAS, March 24, 2009; 106(12): 4584 - 4590.
[Abstract] [Full Text] [PDF]

Home page
 Mol. Biol. CellHome page
Q. Xu, S. Zhu, W. Wang, X. Zhang, W. Old, N. Ahn, and X. Liu
Regulation of Kinetochore Recruitment of Two Essential Mitotic Spindle Checkpoint Proteins by Mps1 Phosphorylation
Mol. Biol. Cell, January 1, 2009; 20(1): 10 - 20.
[Abstract] [Full Text] [PDF]

Home page
 JCBHome page
C. Spycher, E. S. Miller, K. Townsend, L. Pavic, N. A. Morrice, P. Janscak, G. S. Stewart, and M. Stucki
Constitutive phosphorylation of MDC1 physically links the MRE11-RAD50-NBS1 complex to damaged chromatin
J. Cell Biol., April 21, 2008; 181(2): 227 - 240.
[Abstract] [Full Text] [PDF]

Home page
 J. Cell Sci.Home page
C. Richardson, F. H. Rafiqi, H. K. R. Karlsson, N. Moleleki, A. Vandewalle, D. G. Campbell, N. A. Morrice, and D. R. Alessi
Activation of the thiazide-sensitive Na+-Cl- cotransporter by the WNK-regulated kinases SPAK and OSR1
J. Cell Sci., March 1, 2008; 121(5): 675 - 684.
[Abstract] [Full Text] [PDF]

Home page
 J. Biol. Chem.Home page
M. Buxade, N. Morrice, D. L. Krebs, and C. G. Proud
The PSF{middle dot}p54nrb Complex Is a Novel Mnk Substrate That Binds the mRNA for Tumor Necrosis Factor {alpha}
J. Biol. Chem., January 4, 2008; 283(1): 57 - 65.
[Abstract] [Full Text] [PDF]

Home page
 Mol PlantHome page
S. Sullivan, C. E. Thomson, D. J. Lamont, M. A. Jones, and J. M. Christie
In Vivo Phosphorylation Site Mapping and Functional Characterization of Arabidopsis Phototropin 1
Mol Plant, January 1, 2008; 1(1): 178 - 194.
[Abstract] [Full Text] [PDF]

Home page
 Genes Dev.Home page
E. M.J. King, N. Rachidi, N. Morrice, K. G. Hardwick, and M. J.R. Stark
Ipl1p-dependent phosphorylation of Mad3p is required for the spindle checkpoint response to lack of tension at kinetochores
Genes & Dev., May 15, 2007; 21(10): 1163 - 1168.
[Abstract] [Full Text] [PDF]

Home page
 Mol. Cell. Biol.Home page
P. Douglas, X. Cui, W. D. Block, Y. Yu, S. Gupta, Q. Ding, R. Ye, N. Morrice, S. P. Lees-Miller, and K. Meek
The DNA-Dependent Protein Kinase Catalytic Subunit Is Phosphorylated In Vivo on Threonine 3950, a Highly Conserved Amino Acid in the Protein Kinase Domain
Mol. Cell. Biol., March 1, 2007; 27(5): 1581 - 1591.
[Abstract] [Full Text] [PDF]

Home page
 J. Cell Sci.Home page
O. Goransson, M. Deak, S. Wullschleger, N. A. Morrice, A. R. Prescott, and D. R. Alessi
Regulation of the polarity kinases PAR-1/MARK by 14-3-3 interaction and phosphorylation
J. Cell Sci., October 1, 2006; 119(19): 4059 - 4070.
[Abstract] [Full Text] [PDF]


HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS
 All ASBMB Journals   Journal of Biological Chemistry 
 Journal of Lipid Research   ASBMB Today 
Copyright © 2006 by the American Society for Biochemistry and Molecular Biology.
Advertisement
spacer
Advertisement
Advertisement