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Citation
Klement, E., Lipinszki, Z., Kupih├ír, Z., Udvardy, A., Medzihradszky, K.F. (2010). Enrichment of O-GlcNAc modified proteins by the periodate oxidation-hydrazide resin capture approach.  J. Proteome Res. 9(5): 2200--2206.
FlyBase ID
FBrf0210743
Publication Type
Research paper
Abstract
A chemical derivatization approach has been developed for the enrichment of O-GlcNAc modified proteins. The procedure is based on the isolation technique used for N-glycoproteins with appropriate modifications because of the differences in the two types of glycosylation: a prolonged periodate oxidation is followed by hydrazide resin capture, on-resin proteolytic digestion, and release of the modified peptides by hydroxylamine. This enrichment strategy offers a fringe benefit in mass spectrometry analysis. Upon collisional activation, the presence of the open carbohydrate ring leads to characteristic fragmentation facilitating both glycopeptide identification and site assignment. The enrichment protocol was applied to the Drosophila proteasome complex previously described as O-GlcNAc modified. The O-GlcNAc modification was located on proteasome interacting proteins, deubiquitinating enzyme Faf (CG1945) and a ubiquitin-like domain containing protein (CG7546). Three other proteins were also found GlcNAc modified, a HSP70 homologue (CG2918), scribbled (CG5462) and the 205 kDa microtubule-associated protein (CG1483). Interestingly, in the HSP70 homologue the GlcNAc modification is attached to an asparagine residue of a N-glycosylation motif.
PubMed ID
PubMed Central ID
PMC2866058 (PMC) (EuropePMC)
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Secondary IDs
    Language of Publication
    English
    Additional Languages of Abstract
    Parent Publication
    Publication Type
    Journal
    Abbreviation
    J. Proteome Res.
    Title
    Journal of Proteome Research
    Publication Year
    2002
    ISBN/ISSN
    1535-3893
    Data From Reference
    Genes (5)