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Accepted (AM)Open Access (License Unspecified), Open
Journal article
Direct in-gel fluorescence detection and cellular imaging of O-GlcNAc-modified proteins
Journal of the American Chemical Society, v 130(35), pp 11576-11577
03 Sep 2008
PMID: 18683930
Featured in Collection : UN Sustainable Development Goals @ Drexel
Abstract
We report an advanced chemoenzymatic strategy for the direct fluorescence detection, proteomic analysis, and cellular imaging of O-GlcNAc-modified proteins. O-GlcNAc residues are selectively labeled with fluorescent or biotin tags using an engineered galactosyltransferase enzyme and [3 + 2] azide -alkyne cycloaddition chemistry. We demonstrate that this approach can be used for direct in-gel detection and mass spectrometric identification of O-GlcNAc proteins, identifying 146 novel glycoproteins from the mammalian brain. Furthermore, we show that the method can be exploited to quantity dynamic changes in cellular O-GlcNAc levels and to image O-GlcNAc-glycosylated proteins within cells. As such, this strategy enables studies of O-GlcNAc glycosylation that were previously inaccessible and provides a new tool for uncovering the physiological functions of O-GlcNAc.
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Details
- Title
- Direct in-gel fluorescence detection and cellular imaging of O-GlcNAc-modified proteins
- Creators
- Peter M. Clark - California Institute of TechnologyJessica F. Dweck - Genomics Institute of the Novartis Research FoundationDaniel E. Mason - Genomics Institute of the Novartis Research FoundationCourtenay R. Hart - Genomics Institute of the Novartis Research FoundationSuzanne B. Buck - Genomics Institute of the Novartis Research FoundationEric C. Peters - Genomics Institute of the Novartis Research FoundationBrian J. Agnew - Genomics Institute of the Novartis Research FoundationLinda C. Hsieh-Wilson - Genomics Institute of the Novartis Research Foundation
- Publication Details
- Journal of the American Chemical Society, v 130(35), pp 11576-11577
- Publisher
- American Chemical Society; Washington, DC
- Number of pages
- 3
- Grant note
- Howard Hughes Medical Institute RO1 GM084724 / NIH; United States Department of Health & Human Services; National Institutes of Health (NIH) - USA R01GM084724 / NATIONAL INSTITUTE OF GENERAL MEDICAL SCIENCES; United States Department of Health & Human Services; National Institutes of Health (NIH) - USA; NIH National Institute of General Medical Sciences (NIGMS)
- Resource Type
- Journal article
- Language
- English
- Academic Unit
- School of Biomedical Engineering, Science, and Health Systems; Drexel University
- Web of Science ID
- WOS:000258792000007
- Scopus ID
- 2-s2.0-51349166164
- Other Identifier
- 991019356495604721
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- Collaboration types
- Industry collaboration
- Domestic collaboration
- Web of Science research areas
- Chemistry, Multidisciplinary