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Mechanism-Based Probing, Characterization, and Inhibitor Design of Glycosidases and Glycosyltransferases

[ Vol. 9 , Issue. 1 ]


Hiroshi Hinou and Shin-Ichiro Nishimura   Pages 106 - 116 ( 11 )


Recent structural and kinetic studies indicated that enzymes shift their peripheral structures of the catalytic sites dynamically to modify the substrate structures. Molecules which disturb such mechanism of specific enzymes may become potent candidates for therapeutic reagent. This article describes a versatile strategy to synthesize new class of mechanism-based inhibitors for glycosyltransferases and glycoside hydrolases. Combination of irreversible tagging and proteomic analysis of crucial amino acid residues using MALDI-TOF/TOF mass spectrometry allowed a promising method to probe such invisible transitional state in the enzymatic reactions. Feasibility of the fluorescence energy resonance transfer (FRET) is also documented as novel protocol for the real-time and continuous monitoring of glycosyltransferase catalyzed reactions. It was demonstrated that FRET method greatly facilitates discovery research of selective inhibitors in combination with click chemistry.

, Mechanism-Based, Probing,, Characterization,, and, Inhibitor, Design, of, Glycosidases, and, Glycosyltransferases


Graduate School of Advanced Life Science and Frontier Research Center for Post-Genome Science and Technology, Hokkaido University, N21, W11, Sapporo 001-0021, Japan.

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