RESUMEN

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.

Asunto(s)

Ciclohexanoles/farmacología , Disacáridos/farmacología , Compuestos Epoxi/farmacología , alfa-Amilasas Pancreáticas/química , alfa-Amilasas Pancreáticas/metabolismo , Dominio Catalítico , Simulación por Computador , Humanos , Enlace de Hidrógeno , Inositol/análogos & derivados , Inositol/química , Inositol/metabolismo , Cinética , Espectrometría de Masas , Modelos Moleculares , Agua , Difracción de Rayos X