Glycosidase-substrate interactions analysis by STD-NMR spectroscopy: study of alpha-L-fucosidase.
Saturation transfer difference-nuclear magnetic resonance (STD-NMR) is a recently developed method used to study the interaction between large proteins and small ligands. It has been successfully employed for various interactions including those between oligosaccharides or glycomimetics, and binding proteins such as lectins and antibodies. We have demonstrated that this method can be used to directly study the interaction between glycosidases and their substrates. We chose to study alpha-l-fucosidases, which, despite their wide distribution among living organisms and their biological significance, have not been studied with regard to their reaction mechanism. We were able for the first time to show direct evidences for the minimum structural requirements for a compound to interact with these enzymes. These findings will be useful for the design of new inhibitors and to optimize the synthetic properties (transglycosylation) of alpha-l-fucosidases.
Berteau
Olivier
O
Department of Chemistry, Swedish University of Agricultural Sciences, P.O. Box 7015, SE-750 07 Uppsala, Sweden. Olivier.Berteau@kemi.slu.se
Sandström
Corine
C
Bielicki
Julie
J
Anson
Donald S
DS
Kenne
Lennart
L
D017485:1-Deoxynojirimycin / Q000031:analogs & derivatives; D000818:Animals; D004285:Dogs; D004791:Enzyme Inhibitors / Q000737:chemistry / Q000378:metabolism / Q000494:pharmacology; D005643:Fucose / Q000737:chemistry / Q000378:metabolism / Q000494:pharmacology; D050112:Imino Pyranoses; D007700:Kinetics; D019906:Nuclear Magnetic Resonance, Biomolecular / Q000379:methods; D013379:Substrate Specificity; D013402:Sugar Alcohols / Q000737:chemistry / Q000378:metabolism / Q000494:pharmacology; D005644:alpha-L-Fucosidase / Q000037:antagonists & inhibitors / Q000737:chemistry* / Q000378:metabolism
D004791:Enzyme Inhibitors; D050112:Imino Pyranoses; D013402:Sugar Alcohols; D017485:1-Deoxynojirimycin; D005643:Fucose; C062662:deoxyfuconojirimycin; D005644:alpha-L-Fucosidase