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Probing the dynamics of O-GlcNAc glycosylation in the brain using quantitative proteomics

Khidekel, Nelly and Ficarro, Scott B. and Clark, Peter M. and Bryan, Marian C. and Swaney, Danielle L. and Rexach, Jessica E. and Sun, Yi E. and Coon, Joshua J. and Peters, Eric C. and Hsieh-Wilson, Linda C. (2007) Probing the dynamics of O-GlcNAc glycosylation in the brain using quantitative proteomics. Nature Chemical Biology, 3 (6). pp. 339-348. ISSN 1552-4450. http://resolver.caltech.edu/CaltechAUTHORS:20150408-095425960

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Abstract

The addition of the monosaccharide beta-N-acetyl-D-glucosamine to proteins (O-GlcNAc glycosylation) is an intracellular, post-translational modification that shares features with phosphorylation. Understanding the cellular mechanisms and signaling pathways that regulate O-GlcNAc glycosylation has been challenging because of the difficulty of detecting and quantifying the modification. Here, we describe a new strategy for monitoring the dynamics of O-GlcNAc glycosylation using quantitative mass spectrometry-based proteomics. Our method, which we have termed quantitative isotopic and chemoenzymatic tagging (QUIC-Tag), combines selective, chemoenzymatic tagging of O-GlcNAc proteins with an efficient isotopic labeling strategy. Using the method, we detect changes in O-GlcNAc glycosylation on several proteins involved in the regulation of transcription and mRNA translocation. We also provide the first evidence that O-GlcNAc glycosylation is dynamically modulated by excitatory stimulation of the brain in vivo. Finally, we use electron-transfer dissociation mass spectrometry to identify exact sites of O-GlcNAc modification. Together, our studies suggest that O-GlcNAc glycosylation occurs reversibly in neurons and, akin to phosphorylation, may have important roles in mediating the communication between neurons.


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http://dx.doi.org/10.1038/nchembio881DOIArticle
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http://dx.doi.org/10.1038/nchembio0607-303Featured InNature Chemical Biology: News and Views
http://dx.doi.org/10.1038/447510aFeatured InNature: Research Highlights
http://rdcu.be/cZTqFeatured InFree ReadCube access - Nature: Research Highlights
ORCID:
AuthorORCID
Hsieh-Wilson, Linda C.0000-0001-5661-1714
Additional Information:© 2007 Nature Publishing Group. Received 21 December 2006; accepted 13 April 2007; published online 13 May 2007. We thank P. Qasba and B. Ramakrishnan for the generous gift of the GalT plasmid, S. Whiteheart for the OGA antibody, T.C. Neo for assistance with synthesis of ketogalactose probe Compound 1Compound 1, and A. Su for technical discussions. This work was supported by the US National Institutes of Health (RO1 NS045061), the National Science Foundation CAREER Award (CHE-0239861) and the Parson's Foundation (N.K.).
Funders:
Funding AgencyGrant Number
NIHRO1 NS045061
NSFCHE-0239861
Ralph M. Parsons FoundationUNSPECIFIED
Record Number:CaltechAUTHORS:20150408-095425960
Persistent URL:http://resolver.caltech.edu/CaltechAUTHORS:20150408-095425960
Official Citation:Khidekel, N., Ficarro, S. B., Clark, P. M., Bryan, M. C., Swaney, D. L., Rexach, J. E., . . . Hsieh-Wilson, L. C. (2007). Probing the dynamics of O-GlcNAc glycosylation in the brain using quantitative proteomics. [10.1038/nchembio881]. Nat Chem Biol, 3(6), 339-348.
Usage Policy:No commercial reproduction, distribution, display or performance rights in this work are provided.
ID Code:56470
Collection:CaltechAUTHORS
Deposited By: Jason Perez
Deposited On:08 Apr 2015 22:32
Last Modified:07 Nov 2017 22:28

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