Spiro-epoxyglycosides as Activity-Based Probes for Glycoside Hydrolase Family 99 Endomannosidase/Endomannanase.

Autor: Schröder SP; Department of Bioorganic Chemistry, Leiden Institute of Chemistry, Einsteinweg 55, 2333 CC, Leiden, The Netherlands., Kallemeijn WW; Department of Medical Biochemistry, Leiden Institute of Chemistry, Einsteinweg 55, 2333 CC, Leiden, The Netherlands., Debets MF; Department of Bioorganic Chemistry, Leiden Institute of Chemistry, Einsteinweg 55, 2333 CC, Leiden, The Netherlands., Hansen T; Department of Bioorganic Chemistry, Leiden Institute of Chemistry, Einsteinweg 55, 2333 CC, Leiden, The Netherlands., Sobala LF; Department of Chemistry, York Structural Biology Laboratory, University of York, Heslington, York, YO10 5DD, UK., Hakki Z; School of Chemistry and Bio21 Molecular Science and Biotechnology Institute, University of Melbourne, Parkville, Victoria, Australia., Williams SJ; School of Chemistry and Bio21 Molecular Science and Biotechnology Institute, University of Melbourne, Parkville, Victoria, Australia., Beenakker TJM; Department of Bioorganic Chemistry, Leiden Institute of Chemistry, Einsteinweg 55, 2333 CC, Leiden, The Netherlands., Aerts JMFG; Department of Medical Biochemistry, Leiden Institute of Chemistry, Einsteinweg 55, 2333 CC, Leiden, The Netherlands., van der Marel GA; Department of Bioorganic Chemistry, Leiden Institute of Chemistry, Einsteinweg 55, 2333 CC, Leiden, The Netherlands., Codée JDC; Department of Bioorganic Chemistry, Leiden Institute of Chemistry, Einsteinweg 55, 2333 CC, Leiden, The Netherlands., Davies GJ; Department of Chemistry, York Structural Biology Laboratory, University of York, Heslington, York, YO10 5DD, UK., Overkleeft HS; Department of Bioorganic Chemistry, Leiden Institute of Chemistry, Einsteinweg 55, 2333 CC, Leiden, The Netherlands.
Jazyk: angličtina
Zdroj: Chemistry (Weinheim an der Bergstrasse, Germany) [Chemistry] 2018 Jul 11; Vol. 24 (39), pp. 9983-9992. Date of Electronic Publication: 2018 Jun 21.
DOI: 10.1002/chem.201801902
Abstrakt: N-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.
(© 2018 The Authors. Published by Wiley-VCH Verlag GmbH & Co. KGaA.)
Databáze: MEDLINE
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