Title | Glucosyl epi-cyclophellitol allows mechanism-based inactivation and structural analysis of human pancreatic alpha-amylase |
Publication Type | Journal Article |
Year of Publication | 2016 |
Authors | Caner, S, Zhang, X, Jiang, J, Chen, H-M, Nguyen, NT, Overkleeft, H, Brayer, GD, Withers, SG |
Journal | FEBS LETTERS |
Volume | 590 |
Pagination | 1143-1151 |
Date Published | APR |
ISSN | 0014-5793 |
Abstract | As part of a search for selective, mechanism-based covalent inhibitors of human pancreatic alpha-amylase we describe the chemoenzymatic synthesis of the disaccharide analog alpha-glucosyl epi-cyclophellitol, demonstrate its stoichiometric reaction with human pancreatic alpha-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. |
DOI | 10.1002/1873-3468.12143 |
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