Photoactivatable Glycopolymers for the Proteome-Wide Identification of Fucose-α(1-2)-Galactose Binding Proteins
Abstract
Although fucose-α(1-2)-galactose (Fucα(1-2)Gal)-containing glycans have been implicated in cognitive processes such as learning and memory, their precise molecular mechanisms are poorly understood. Here we employed the use of multivalent glycopolymers to enable the first proteome-wide identification of weak affinity, low abundance Fucα(1-2)Gal glycan-binding proteins (GBPs). Biotin-terminated glycopolymers containing photoactivatable cross-linking groups were designed to capture and enrich GBPs from rat brain lysates. Candidate proteins were tested for their ability to bind Fucα(1-2)Gal, and the functional significance of the interaction was investigated for the synaptic vesicle protein SV2a using a knockout mouse system. The results suggest a role for SV2a-Fucα(1-2)Gal interactions in SV2a trafficking and synaptic vesicle recycling. More broadly, our studies outline a general chemical approach for the systems-level discovery of novel GBPs.
Additional Information
© 2014 American Chemical Society. Received: March 11, 2014. Publication Date (Web): June 17, 2014. ACS AuthorChoice Open Access article. We thank Drs. J. Lowe and S. Domino for the FUT KO mice, Dr. S. Bajjalieh for the SV2a plasmid, Dr. J. Gleeson for the DCLK1 plasmid, Dr. T. Kosaza for GαO protein, Dr. M. Shahgoli in the CCE Division Mass Spectrometry Facility, and Dr. C. Krishnamurthy for BgtA enzyme. This work was supported by the NIH (R01 GM084724).Attached Files
Published - ja502482a.pdf
Supplemental Material - ja502482a_si_001.pdf
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Additional details
- PMCID
- PMC4105059
- Eprint ID
- 48633
- Resolver ID
- CaltechAUTHORS:20140815-153145479
- R01 GM084724
- NIH
- Created
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2014-08-19Created from EPrint's datestamp field
- Updated
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2021-11-10Created from EPrint's last_modified field