Powerful probes for glycosidases: novel, fluorescently tagged glycosidase inhibitors

• Published in: Bioorganic & medicinal chemistry letters Vol. 11; no. 10; pp. 1339 - 1342
• Main Authors: Hermetter, Albin, Scholze, Hubert, Stütz, Arnold E., Withers, Stephen G., Wrodnigg, Tanja M.
• Format: Journal Article
• Language: English
• Published: England Elsevier Ltd 21.05.2001
• Subjects: Binding, Competitive; Dansyl Compounds - chemistry; Dose-Response Relationship, Drug; Electrophoresis, Polyacrylamide Gel - methods; Enzyme Inhibitors - chemical synthesis; Enzyme Inhibitors - metabolism; Fluorescent Dyes - chemical synthesis; Fluorescent Dyes - metabolism; Glycoside Hydrolases - antagonists & inhibitors; Mannitol - chemical synthesis; Mannitol - metabolism; Molecular Probes - chemical synthesis; Molecular Probes - metabolism; Protein Binding; Rhizobium - enzymology
• ISSN: 0960-894X; 1464-3405
• DOI: 10.1016/S0960-894X(01)00209-8

1-Amino-1,2,5-trideoxy-2,5-imino- d-mannitol was fluorescently tagged by reaction with dansyl chloride at N-1 or by attachment of a dansyl amide bearing spacer to this centre. Compounds obtained are highly potent inhibitors of β-glucosidase exhibiting K i values in the single figure nanomolar range. The 1- N-dansyl substituted inhibitor was successfully exploited for binding studies with β-glucosidase from Agrobacterium sp. employing fluorescence spectrometric methods. 1- N-Dansyl derivatives of 1-amino-1,2,5-trideoxy-2,5-imino- d-mannitol exhibiting K i values in the low nanomolar range are reported. An enzyme/inhibitor complex with Agrobacterium sp. β-glucosidase was characterised by fluorescence spectrometry.