5EMY

Human Pancreatic Alpha-Amylase in complex with the mechanism based inactivator glucosyl epi-cyclophellitol

Summary for 5EMY

• Entry DOI: 10.2210/pdb5emy/pdb
• Related: 1CPU 4W93 4X9Y
• Descriptor: Pancreatic alpha-amylase, (1R,2R,3S,5R,6S)-2,3,5-trihydroxy-6-(hydroxymethyl)cyclohexyl alpha-D-glucopyranoside, CHLORIDE ION, ... (5 entities in total)
• Functional Keywords: amylase, diabetes, obesity, glucosyl hydrolase, hydrolase-hydrolase inhibitor complex, hydrolase/hydrolase inhibitor
• Biological source: Homo sapiens (Human)
• Total number of polymer chains: 1
• Total formula weight: 56347.16

Authors

Caner, S.,Brayer, G.D. (deposition date: 2015-11-06, release date: 2016-07-06, Last modification date: 2024-10-23)

Primary citation

Caner, S.,Zhang, X.,Jiang, J.,Chen, H.M.,Nguyen, N.T.,Overkleeft, H.,Brayer, G.D.,Withers, S.G.
Glucosyl epi-cyclophellitol allows mechanism-based inactivation and structural analysis of human pancreatic alpha-amylase.
Febs Lett., 590:1143-1151, 2016

PubMed Abstract: 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.

PubMed: 27000970
DOI: 10.1002/1873-3468.12143

Experimental method

X-RAY DIFFRACTION (1.231 Å)