title: Synthesis and analysis of specific covalent inhibitors of endo-xyloglucanases.
abstract: A series of N-bromoacetylglycosylamines and bromoketone C-glycosides were synthesised from complex xyloglucan oligosaccharide (XyGO) scaffolds as specific active-site affinity labels for endo-xyloglucanases. Compounds based on XXXG (Xyl3 Glc4 ) and XLLG (Xyl3 Glc4 Gal2 ) oligosaccharides exhibited strikingly higher affinities and higher rates of irreversible inhibition than known cellobiosyl and new lactosyl disaccharide congeners when tested with endo-xyloglucanases from two distinct glycoside hydrolase (GH) families. Intact-protein mass spectrometry indicated that inactivation with XyGO derivatives generally resulted in a 1:1 labelling stoichiometry. Together, these results indicate that XyGO-based affinity reagents have significant potential as inhibitors and proteomic reagents for the identification and analysis of diverse xyloglucan-active enzymes in nature, to facilitate industrial enzyme applications.
authors: Michael Smith Laboratories and Department of Chemistry, University of British Columbia, 2185 East Mall, Vancouver, British Columbia V6T 1Z4 (Canada).
authors: Thomas Hauch
authors: TH
authors: Fenger
authors: Harry
authors: H
authors: Brumer
mesh_terms: D001439:Bacteroides / Q000201:enzymology*; D002240:Carbohydrate Sequence; D048013:Acetivibrio thermocellus / Q000201:enzymology*; D004791:Enzyme Inhibitors / Q000737:chemistry / Q000494:pharmacology; D005936:Glucans / Q000378:metabolism; D006026:Glycoside Hydrolases / Q000037:antagonists & inhibitors* / Q000378:metabolism; D008969:Molecular Sequence Data; D009844:Oligosaccharides / Q000737:chemistry* / Q000494:pharmacology*; D013379:Substrate Specificity; D014990:Xylans / Q000378:metabolism
chemical_list: D004791:Enzyme Inhibitors; D005936:Glucans; D009844:Oligosaccharides; D014990:Xylans; C029353:xyloglucan; D006026:Glycoside Hydrolases; C072818:xyloglucan endo(1-4)-beta-D-glucanase
keywords: carbohydrates; enzymes; glycosyl hydrolases; inhibitors; xyloglucanases