Glucosyl epi-cyclophellitol allows mechanism-based inactivation and structural analysis of human pancreatic α-amylase
| Autor: | Xiaohua Zhang, Sami Caner, Gary D. Brayer, Jianbing Jiang, Stephen G. Withers, Nham T. Nguyen, Hermen S. Overkleeft, Hong-Ming Chen |
|---|---|
| Rok vydání: | 2015 |
| Předmět: |
glycosyl enzyme
Models Molecular amylase Stereochemistry activity-based probes Kinetics Biophysics Disaccharide Pancreatic alpha-Amylases 010402 general chemistry Disaccharides 01 natural sciences Biochemistry conduritol epoxide Mass Spectrometry chemistry.chemical_compound Nucleophile X-Ray Diffraction Structural Biology GH13 structure Catalytic Domain Genetics Humans Computer Simulation Amylase Molecular Biology mechanism-based inhibition biology 010405 organic chemistry Hydrogen bond Active site Water Hydrogen Bonding Cell Biology Cyclohexanols 0104 chemical sciences chemistry Covalent bond biology.protein Epoxy Compounds Derivative (chemistry) Inositol |
| Zdroj: | FEBS letters. 590(8) |
| ISSN: | 1873-3468 |
| Popis: | As part of a search for selective, mechanism-based covalent inhibitors of human pancreatic α-amylase we describe the chemoenzymatic synthesis of the disaccharide analog α-glucosyl epi-cyclophellitol, demonstrate its stoichiometric reaction with human pancreatic α-amylase and evaluate the time dependence of its inhibition. X-ray crystallographic analysis of the covalent derivative so formed confirms its reaction at the active site with formation of a covalent bond to the catalytic nucleophile D197. The structure illuminates the interactions with the active site and confirms OH4' on the nonreducing end sugar as a good site for attachment of fluorescent tags in generating probes for localization and quantitation of amylase in vivo. |
| Databáze: | OpenAIRE |
| Externí odkaz: |
|
Plný text