Fluorescent-tagged sp2-iminosugars with potent β-glucosidase inhibitory activity.
New fluorescently-labelled sp(2)-iminosugars based on the 5N,6S-[N'-(4-aminobutyl)iminomethylidene]-6-thionojirimycin skeleton have been synthesized as photoprobes to monitor glycosidase binding. Dansyl, dapoxyl and coumarin fluorophores were appended to the terminal amino group at the N'-substituent by either sulfonamide or amide bridging reaction. All the conjugates behaved as strong (low micromolar to nanomolar) and selective inhibitors of β-glucosidases (almonds and bovine liver) and naringinase, in agreement with the inhibition pattern previously encountered for related iso(thio)urea-type bicyclic sp(2)-iminosugars. The presence of the fluorescent probe allows real-time and continuous monitoring of β-glucosidase inhibition by fluorescence resonance energy transfer (FRET), taking advantage of the intrinsic tryptophan-associated fluorescence of the protein.
Aguilar-Moncayo
Matilde
M
Departamento de Química Orgánica, Facultad de Química, Universidad de Sevilla, Apartado 553, E-41012 Sevilla, Spain.
García-Moreno
M Isabel
MI
Stütz
Arnold E
AE
García Fernández
José M
JM
Wrodnigg
Tanja M
TM
Ortiz Mellet
Carmen
C
D017485:1-Deoxynojirimycin / Q000031:analogs & derivatives / Q000737:chemistry; D000818:Animals; D002417:Cattle; D004791:Enzyme Inhibitors / Q000138:chemical synthesis / Q000737:chemistry* / Q000494:pharmacology; D031541:Fluorescence Resonance Energy Transfer; D005456:Fluorescent Dyes / Q000737:chemistry*; D050111:Imino Sugars / Q000737:chemistry*; D009097:Multienzyme Complexes / Q000037:antagonists & inhibitors / Q000378:metabolism; D001617:beta-Glucosidase / Q000037:antagonists & inhibitors* / Q000378:metabolism
D004791:Enzyme Inhibitors; D005456:Fluorescent Dyes; D050111:Imino Sugars; D009097:Multienzyme Complexes; D017485:1-Deoxynojirimycin; C039827:naringinase; D001617:beta-Glucosidase; C013626:nojirimycin