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dc.contributor.authorSchroder, SP
dc.contributor.authorKallemeijn, WW
dc.contributor.authorDebets, MF
dc.contributor.authorHansen, T
dc.contributor.authorSobala, LF
dc.contributor.authorHakki, Z
dc.contributor.authorWilliams, SJ
dc.contributor.authorBeenakker, TJM
dc.contributor.authorAerts, JMFG
dc.contributor.authorvan der Marel, GA
dc.contributor.authorCodee, JDC
dc.contributor.authorDavies, GJ
dc.contributor.authorOverkleeft, HS
dc.date.accessioned2020-12-17T04:20:23Z
dc.date.available2020-12-17T04:20:23Z
dc.date.issued2018-07-11
dc.identifier.citationSchroder, S. P., Kallemeijn, W. W., Debets, M. F., Hansen, T., Sobala, L. F., Hakki, Z., Williams, S. J., Beenakker, T. J. M., Aerts, J. M. F. G., van der Marel, G. A., Codee, J. D. C., Davies, G. J. & Overkleeft, H. S. (2018). Spiro-epoxyglycosides as Activity-Based Probes for Glycoside Hydrolase Family 99 Endomannosidase/Endomannanase. CHEMISTRY-A EUROPEAN JOURNAL, 24 (39), pp.9983-9992. https://doi.org/10.1002/chem.201801902.
dc.identifier.issn0947-6539
dc.identifier.urihttp://hdl.handle.net/11343/255266
dc.description.abstractN-Glycans direct protein function, stability, folding and targeting, and influence immunogenicity. While most glycosidases that process N-glycans cleave a single sugar residue at a time, enzymes from glycoside hydrolase family 99 are endo-acting enzymes that cleave within complex N-glycans. Eukaryotic Golgi endo-1,2-α-mannosidase cleaves glucose-substituted mannose within immature glucosylated high-mannose N-glycans in the secretory pathway. Certain bacteria within the human gut microbiota produce endo-1,2-α-mannanase, which cleaves related structures within fungal mannan, as part of nutrient acquisition. An unconventional mechanism of catalysis was proposed for enzymes of this family, hinted at by crystal structures of imino/azasugars complexed within the active site. Based on this mechanism, we developed the synthesis of two glycosides bearing a spiro-epoxide at C-2 as electrophilic trap, to covalently bind a mechanistically important, conserved GH99 catalytic residue. The spiro-epoxyglycosides are equipped with a fluorescent tag, and following incubation with recombinant enzyme, allow concentration, time and pH dependent visualization of the bound enzyme using gel electrophoresis.
dc.languageEnglish
dc.publisherWILEY-V C H VERLAG GMBH
dc.titleSpiro-epoxyglycosides as Activity-Based Probes for Glycoside Hydrolase Family 99 Endomannosidase/Endomannanase
dc.typeJournal Article
dc.identifier.doi10.1002/chem.201801902
melbourne.affiliation.departmentSchool of Chemistry
melbourne.source.titleChemistry: A European Journal
melbourne.source.volume24
melbourne.source.issue39
melbourne.source.pages9983-9992
dc.rights.licenseCC BY-NC-ND
melbourne.elementsid1337765
melbourne.contributor.authorWilliams, Spencer
dc.identifier.eissn1521-3765
melbourne.accessrightsOpen Access


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