title: 2-Deoxy-2-fluoro-D-glycosyl fluorides. A new class of specific mechanism-based glycosidase inhibitors.
abstract: Mechanism-based glycosidase inhibitors are of considerable use in studies of enzyme mechanism, in studies of glycoprotein processing, and possibly therapeutically in control of sugar uptake. This paper describes a new general approach to mechanism-based inactivation of glycosidases which involves trapping a covalent glycosyl enzyme intermediate. This is achieved by use of 2-deoxy-2-fluoro-D-glycosyl fluorides, for which the rate of hydrolysis of the fluoroglycosyl enzyme intermediate is extremely slow, resulting in accumulation of the intermediate. Eleven different glycosidases were tested with their corresponding 2-deoxy-2-fluoro-D-glycosyl fluorides. Eight of the eleven were inactivated, four of them according to pseudo first-order kinetics and four according to a more complex kinetic scheme. The specificity of these inhibitors was investigated by assaying for inhibition of one enzyme with four different 2-deoxy-2-fluoro-D-glycosyl fluorides. Large differences in inactivation rate were observed which paralleled previously observed substrate specificities.
authors.affiliation: Department of Chemistry, University of British Columbia, Vancouver, Canada.
authors.forename: S G
authors.initials: SG
authors.lastname: Withers
authors.forename: K
authors.initials: K
authors.lastname: Rupitz
authors.forename: I P
authors.initials: IP
authors.lastname: Street
mesh_terms: D000421:Alcaligenes / Q000201:enzymology; D000818:Animals; D002417:Cattle; D004926:Escherichia coli / Q000201:enzymology; D005461:Fluorine / Q000494:pharmacology*; D006026:Glycoside Hydrolases / Q000037:antagonists & inhibitors*; D006027:Glycosides / Q000494:pharmacology*; D008956:Models, Chemical; D010944:Plants / Q000201:enzymology; D012441:Saccharomyces cerevisiae / Q000201:enzymology
chemical_list: D006027:Glycosides; D005461:Fluorine; D006026:Glycoside Hydrolases