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


     


Originally published In Press as doi:10.1074/mcp.T500040-MCP200 on February 1, 2006.
This Article
Right arrow Full Text
Right arrow Full Text (PDF)
Right arrow Supplemental Data
Right arrow All Versions of this Article:
T500040-MCP200v1
5/5/923    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 Vosseller, K.
Right arrow Articles by Burlingame, A. L.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Vosseller, K.
Right arrow Articles by Burlingame, A. L.
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 5:923-934, 2006.
© 2006 by The American Society for Biochemistry and Molecular Biology, Inc.


Technology

O-Linked N-Acetylglucosamine Proteomics of Postsynaptic Density Preparations Using Lectin Weak Affinity Chromatography and Mass Spectrometry*,S

Keith Vosseller{ddagger},§, Jonathan C. Trinidad{ddagger}, Robert J. Chalkley{ddagger}, Christian G. Specht, Agnes Thalhammer, Aenoch J. Lynn{ddagger}, June O. Snedecor{ddagger}, Shenheng Guan{ddagger}, Katalin F. Medzihradszky{ddagger}, David A. Maltby{ddagger}, Ralf Schoepfer and Alma L. Burlingame{ddagger},||

From the {ddagger} Mass Spectrometry Facility, Department of Pharmaceutical Chemistry, University of California, San Francisco, California 94143 and Laboratory for Molecular Pharmacology, Department of Pharmacology, University College London, London WC1E 6BT, United Kingdom

O-GlcNAc is a widespread dynamic carbohydrate modification of cytosolic and nuclear proteins with features analogous to phosphorylation. O-GlcNAc acts critically in many cellular processes, including signal transduction, protein degradation, and regulation of gene expression. However, the study of its specific regulatory functions has been limited by difficulties in mapping sites of O-GlcNAc modification. We report methods for direct enrichment and identification of in vivo O-GlcNAc-modified peptides through lectin weak affinity chromatography (LWAC) and mass spectrometry. The effectiveness of this strategy on complex peptide mixtures was demonstrated through enrichment of 145 unique O-GlcNAc-modified peptides from a postsynaptic density preparation. 65 of these O-GlcNAc-modified peptides were sequenced and belonged to proteins with diverse functions in synaptic transmission. ß-Elimination/Michael addition, MS3 on O-GlcNAc neutral loss ions, and electron capture dissociation were shown to facilitate analysis of O-GlcNAc-modified peptides/sites from lectin weak affinity chromatography enriched postsynaptic density samples. Bassoon and Piccolo, proteins critical to synapse assembly and vesicle docking, were extensively modified by O-GlcNAc. In some cases, O-GlcNAc was mapped to peptides previously identified as phosphorylated, indicating potential interplay between these modifications. Shared substrate amino acid context was apparent in subsets of O-GlcNAc-modified peptides, including "PVST" and a novel "TTA" motif (two hydroxyl-containing amino acids adjacent to an alanine). The results suggest specific roles for O-GlcNAc modification in synaptic transmission, establish a basis for site-specific regulatory studies, and provide methods that will facilitate O-GlcNAc proteome analysis across a wide variety of cells and tissues.


§ To whom correspondence may be addressed: Dept. of Biochemistry and Molecular Biology, Drexel University College of Medicine, 245 N. 15th St., M. S. 497, Philadelphia, PA 19102. Tel.: 215-762-8789; E-mail: kav27{at}drexel.edu


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
Proc. Natl. Acad. Sci. USAHome page
R. J. Chalkley, A. Thalhammer, R. Schoepfer, and A. L. Burlingame
Identification of protein O-GlcNAcylation sites using electron transfer dissociation mass spectrometry on native peptides
PNAS, June 2, 2009; 106(22): 8894 - 8899.
[Abstract] [Full Text] [PDF]


Home page
Brief Funct Genomic ProteomicHome page
X. Wei and L. Li
Comparative glycoproteomics: approaches and applications
Brief Funct Genomic Proteomic, March 1, 2009; 8(2): 104 - 113.
[Abstract] [Full Text] [PDF]


Home page
GlycobiologyHome page
J. F Valliere-Douglass, L. J Brady, C. Farnsworth, D. Pace, A. Balland, A. Wallace, W. Wang, M. J Treuheit, and B. Yan
O-Fucosylation of an antibody light chain: Characterization of a modification occurring on an IgG1 molecule
Glycobiology, February 1, 2009; 19(2): 144 - 152.
[Abstract] [Full Text] [PDF]


Home page
J. Biol. Chem.Home page
M. K. Tallent, N. Varghis, Y. Skorobogatko, L. Hernandez-Cuebas, K. Whelan, D. J. Vocadlo, and K. Vosseller
In Vivo Modulation of O-GlcNAc Levels Regulates Hippocampal Synaptic Plasticity through Interplay with Phosphorylation
J. Biol. Chem., January 2, 2009; 284(1): 174 - 181.
[Abstract] [Full Text] [PDF]


Home page
Am. J. Physiol. Endocrinol. Metab.Home page
R. J. Copeland, J. W. Bullen, and G. W. Hart
Cross-talk between GlcNAcylation and phosphorylation: roles in insulin resistance and glucose toxicity
Am J Physiol Endocrinol Metab, July 1, 2008; 295(1): E17 - E28.
[Abstract] [Full Text] [PDF]


Home page
Mol. Cell. ProteomicsHome page
J. C. Trinidad, A. Thalhammer, C. G. Specht, A. J. Lynn, P. R. Baker, R. Schoepfer, and A. L. Burlingame
Quantitative Analysis of Synaptic Phosphorylation and Protein Expression
Mol. Cell. Proteomics, April 1, 2008; 7(4): 684 - 696.
[Abstract] [Full Text] [PDF]


Home page
Mol. Cell. ProteomicsHome page
Z. Wang, A. Pandey, and G. W. Hart
Dynamic Interplay between O-Linked N-Acetylglucosaminylation and Glycogen Synthase Kinase-3-dependent Phosphorylation
Mol. Cell. Proteomics, August 1, 2007; 6(8): 1365 - 1379.
[Abstract] [Full Text] [PDF]


Home page
Am. J. Physiol. Renal Physiol.Home page
M. G. Janech, J. R. Raymond, and J. M. Arthur
Proteomics in renal research
Am J Physiol Renal Physiol, February 1, 2007; 292(2): F501 - F512.
[Abstract] [Full Text] [PDF]


Home page
Mol. Cell. ProteomicsHome page
J. C. Trinidad, C. G. Specht, A. Thalhammer, R. Schoepfer, and A. L. Burlingame
Comprehensive Identification of Phosphorylation Sites in Postsynaptic Density Preparations
Mol. Cell. Proteomics, May 1, 2006; 5(5): 914 - 922.
[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