Staff Publications

Staff Publications

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    'Staff publications' is the digital repository of Wageningen University & Research

    'Staff publications' contains references to publications authored by Wageningen University staff from 1976 onward.

    Publications authored by the staff of the Research Institutes are available from 1995 onwards.

    Full text documents are added when available. The database is updated daily and currently holds about 240,000 items, of which 72,000 in open access.

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Record number 436275
Title Structural and mechanistic insight into N-glycan processing by endo-a-mannosidase
Author(s) Thompson, A.J.; Williams, R.J.; Hakki, Z.; Alonzi, D.S.; Wennekes, T.; Gloster, T.M.; Songsrirote, K.; Thomas-Oates, E.; Wrodnigg, T.M.; Spreitz, J.; Stütz, A.E.; Butters, T.D.; Williams, S.J.; Davies, G.J.
Source Proceedings of the National Academy of Sciences of the United States of America 109 (2012)3. - ISSN 0027-8424 - p. 781 - 786.
DOI https://doi.org/10.1073/pnas.1111482109
Department(s) Laboratory for Organic Chemistry
VLAG
Publication type Refereed Article in a scientific journal
Publication year 2012
Keyword(s) asparagine-linked oligosaccharides - glucosidase-ii-deficient - glycoprotein-biosynthesis - endomannosidase pathway - endoplasmic-reticulum - glycoside hydrolases - quality-control - inhibitors - cells - classification
Abstract N-linked glycans play key roles in protein folding, stability, and function. Biosynthetic modification of N-linked glycans, within the endoplasmic reticulum, features sequential trimming and readornment steps. One unusual enzyme, endo-a-mannosidase, cleaves mannoside linkages internally within an N-linked glycan chain, short circuiting the classical N-glycan biosynthetic pathway. Here, using two bacterial orthologs, we present the first structural and mechanistic dissection of endo-a-mannosidase. Structures solved at resolutions 1.7–2.1 Å reveal a (ß/a)8 barrel fold in which the catalytic center is present in a long substrate-binding groove, consistent with cleavage within the N-glycan chain. Enzymatic cleavage of authentic Glc1/3Man9GlcNAc2 yields Glc1/3-Man. Using the bespoke substrate a-Glc-1,3-a-Man fluoride, the enzyme was shown to act with retention of anomeric configuration. Complexes with the established endo-a-mannosidase inhibitor a-Glc-1,3-deoxymannonojirimycin and a newly developed inhibitor, a-Glc-1,3-isofagomine, and with the reducing-end product a-1,2-mannobiose structurally define the -2 to +2 subsites of the enzyme. These structural and mechanistic data provide a foundation upon which to develop new enzyme inhibitors targeting the hijacking of N-glycan synthesis in viral disease and cancer.
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